Team:Toulouse INSA-UPS/Materials

Plasmids



  • pAM4951: Cyanobacteria cloning vector used as a backbone for pCONCOMBRE. It is designed for chromosomal integration into Synechococcus elongatus neutral site 1 (NS1). The device has an origin of replication for E. coli derived from pBR322 and an oriT site for conjugal transfer recognized by the RK2/RP4 mobilization machinery. The device confers spectinomycin and streptomycin (SpSm) resistance to both E. coli and S. elongatus. pAM4951 (pCV0063) was a gift from Susan Golden (Addgene plasmid # 120084 ; http://n2t.net/addgene:120084; RRID:Addgene_120084).

  • pRL443: Conjugal plasmid used for triparental conjugation (cyanobacteria transformation). This plasmid contains the many genes required to encode the conjugal apparatus. The device confers Chloramphenicol resistance. pRL443 was a gift from Peter Wolk (Addgene plasmid # 70261 ; http://n2t.net/addgene:70261 ; RRID:Addgene_70261).

  • pRL623: Helper plasmid for bacterial conjugal DNA transfer (cyanobacteria transformation). It encodes for specific DNA-methylases that act on the cargo vector so that it is not degraded after entry into the cyanobacterium cellular space. The device confers Ampicillin resistance. pRL623 was a gift from Peter Wolk (Addgene plasmid # 58494 ; http://n2t.net/addgene:58494 ; RRID:Addgene_58494).

  • pCfB3034: EasyClone-MarkerFree backbone vector for integration into site X-3 (Chr X: 223616..224744) of S. cerevisiae. Used as a backbone for the cloning of pFRAMBOISE. Transformed E. coli cells are resistant to ampicillin. pCfB3034(X-3 MarkerFree) was a gift from Irina Borodina (Addgene plasmid # 73272 ; http://n2t.net/addgene:73272 ; RRID:Addgene_73272).

  • pCfB3038: EasyClone-MarkerFree backbone vector for integration into site XII-1 (Chr XII: 795787..796720) of S. cerevisiae. Used as a backbone for the cloning of pFLEUR. Transformed E. coli cells are resistant to ampicillin. pCfB3038(XII-1 MarkerFree) was a gift from Irina Borodina (Addgene plasmid # 73278 ; http://n2t.net/addgene:73278 ; RRID:Addgene_73278).

  • pCfB3040: EasyClone-MarkerFree backbone vector for integration into site XII-4 (Chr XII:830227..831248) of S. cerevisiae. Used as a backbone for the cloning of pVIOLETTE. Transformed E. coli cells are resistant to ampicillin. pCfB3040(XII-4 MarkerFree) was a gift from Irina Borodina (Addgene plasmid # 73280 ; http://n2t.net/addgene:73280 ; RRID:Addgene_73280).

  • pSH65: Cre-loxP plasmid used to remove the kanMX gene from the LycoYeast strain. Encodes for the Cre-recombinase under the control of the galactose-inducible promoter GAL1 (pGAL1-cre). Transformed E. coli cells are resistant to ampicillin. The marker gene for yeast transformation is pAgTEF1-ble-tScCYC1 and the selectable phenotype is Phleomycin resistance. pSH65 was a gift from Sara Castaño Cerezo (http://www.euroscarf.de/plasmid_details.php?accno=P30122).

  • pATP416-­pluxO5-­ptetO7-­crtYBI: Plasmid used to recover the sequence of the ptetO7 promoter by PCR for use in the construction pFRAMBOISE. Transformed E. coli cells are resistant to ampicillin. pATP416-­pluxO5-­ptetO7-­crtYBI was a gift from Jun Ishii (Addgene plasmid # 165975 ; http://n2t.net/addgene:165975 ; RRID:Addgene_165975).

Strains



E. coli


  • E. coli Stellar: (F–, endA1, supE44, thi-1, recA1, relA1, gyrA96, phoA, Φ80d lacZΔ M15, Δ(lacZYA-argF) U169, Δ(mrr-hsdRMS-mcrBC), ΔmcrA, λ–) used for InFusion cloning.

  • E. coli HB101: (F–, thi-1, hsdS20 (rB–, mB–), supE44, recA13, ara-14, leuB6, proA2, lacY1, galK2, rpsL20 (strr), xyl-5, mtl-1) used for triparental conjugation.


S. cerevisiae


  • LycoYeast: Strain named originally yGPP034 ipENZ011 ipENZ078 based on the parental laboratory strain CEN-PK2-1D (MATalpha; his3D1; leu2-3_112; ura3-52; trp1-289; MAL2-8c; SUC2). This strain produces a quantity of lycopene of the order of 8 mg/gCDW thanks to the modifications listed below:

    • - ΔDPP1: Deletion of the DIACYLGLYCEROL PYROPHOSPHATE PHOSPHATASE 1 which is responsible for the catalysis of the dephosphorylation of diacylglycerol diphosphate (GGPP), the precursor of phytoene, to phosphatidate (PA).

    • - +HMG1: Addition of the 3-HYDROXY-3-METHYLGLUTARYL-COENZYME A REDUCTASE 1 which is responsible for the catalysis of the conversion of HMG-CoA to mevalonate.

    • - +ERG20: Addition of the FARNESYL PYROPHOSPHATE SYNTHASE in charge of the catalysis of the sequential condensation of isopentenyl pyrophosphate with the allylic pyrophosphates, dimethylallyl pyrophosphate, and then with the resultant geranylpyrophosphate to the ultimate product farnesyl pyrophosphate.

    • - +CrtB/E/I: Addition of GERANYLGERANYL DIPHOSPHATE SYNTHASE, 15-CIS-PHYTOENE SYNTHASE AND PHYTOENE DESATURASE (lycopene-forming) which are responsible for the catalysis of the synthese of GGPP, phytoene and lycopene respectively.


S. elongatus


  • WT S. elongatus UTEX 2973: Fast growing cyanobacterial strain discovered in 2015. Fore more information, see (Yu et al. 2015).

  • S. elongatus UTEX 2973 CscB: Wild Type strain engineered to express the sucrose/H+ symporter CscB, from E. coli under the control of the IPTG-inducible promoter PlacUV5. This modification leads to sucrose production with a rate of 1.9 g.L−1day−1 in salt stress conditions, reaching titers as high as 8 g.L−1 (Lin et al. 2020).

  • S. elongatus UTEX 2973 CscB SPS SPP: S. elongatus UTEX 2973 CscB strain in which are also overexpressed the sucrose-phosphate synthase (sps) and sucrose-phosphate phosphatase (spp) genes from Synechocystis sp. PCC 6803 resulting in a sucrose secretion with a rate of 1.4 g.L−1day−1 without additional NaCl (Lin et al. 2020).

Primers



Set of primers used for gblocks amplification


gblocks name

Primer pairs name

5’ to 3’ sequence

Pcyc-neoR-Tcyc

pFRAMBOISEfused_neoR_Forward

ataaacaaacaaagtcgacccagtggtctttcagat

pFRAMBOISEfused_neoR_Reverse

tagtcctgcaggttgtggatgatttataatgttacatgcgtacac

Ptet

pFRAMBOISEfused_Ptet_Forward

cgcatggaatcgtacgtctctatcactgatagggagtaaactcgaca

pFRAMBOISEfused_Ptet_Reverse

ccactgggtcgactttgtttgtttatgtgtgtttattcgaaactaagttct

pCfB3032

pFRAMBOISEfused_iPCRpCfB3032_Forward

aaatcatccacaacctgcaggactagtgctgagg

pFRAMBOISEfused_iPCRpCfB3032_Reverse

gtgatagagacgtacgattccatgcgagctcgctga

pFRAMB

pFRAMBOISEfused_iPCRpFRAMB_Forward

acagaacagcgtcgacccagtggtctttcagat

pFRAMBOISEfused_iPCRpFRAMB_Reverse

tttagcacctaggtttgtttgtttatgtgtgtttattcgaaactaagttct

Tvps13-Tgyp7-rtTA-Phxt7

pFRAMBOISEfused_rtTA_Forward

caaactgtgagctagctcacatatgaaagtatatacccgcttttgtaca

pFRAMBOISEfused_rtTA_Reverse

agaccactgggtcgacgctgttctgtccgt

CrtY-Linker-phCCD1

pFRAMBOISEfused_CrtY-phCCD1_Forward

ataaacaaacaaacctaggtgctaaaaaatgcaaccacatt

pFRAMBOISEfused_CrtY-phCCD1_Reverse

tcatatgtgagctagctcacagtttggcttgttcttgaatttgtt

Linker-phCCD1-Tvps13

pFRAMBOISEnotfused_phCCD1_Forward

agataaggaaaggtcagcaaattctggtg

pFRAMBOISEnotfused_phCCD1_Reverse

tgaagaaaaacgcgctgcggatatttctaag

CrtY-Tadh1-Ptef1-fyn

pFRAMBOISEnotfused_CrtY_Forward

aacaaacaaaatgcaacctcactacgac

pFRAMBOISEnotfused_CrtY_Reverse

ttgctgacctttccttatctttgcattgaac

pFRAMBOI

pFRAMBOISEnotfused_pFRAMBOI_Forward

ccgcagcgcgtttttcttcagtttttccgtttc

pFRAMBOISEnotfused_pFRAMBOI_Reverse

gaggttgcattttgtttgtttatgtgtgtttattc

pCfB3040

pVIOLETTE_iPCRpCfB3040_Forward

gagctcgctgaggacttaattgccata

pVIOLETTE_iPCRpCfB3040_Reverse

tgtaacattataatggcgctctgcaggactagtgctga

Linker-ofCCD1m-Tvps13

pVIOLETTE_ofCCD1m_Forward

tggcaattaagtcctcagcgagctcggcggtggcggaagcggt

pVIOLETTE_ofCCD1m_Reverse

tttgaagctatgcagcgccaccatggcgctgttcgcgct

Pgal-LcyE-Tvps13

pVIOLETTE_LcyE_Forward

tatggcaattaagtcctcagcgagctctaagttggtgtggcggatt

pVIOLETTE_LcyE_Reverse

ccaccgcttccgccaccgccgctgccacctccgccgct

Ptef1-nsrR-Tcyc1

pVIOLETTE_nsrR_Forward

tatccgcagcgcgaacagcgccatggtggcgctgcata

pVIOLETTE_nsrR_Reverse

ttaatgcctcagcactagtcctgcagagcgccattata

Final Round

pVIOLETTE_FR_Forward

actggtctctttagagggaactaggacattcttt

pVIOLETTE_FR_Reverse

ttaccagagatgccatatttcggcgc

New-LcyE

pVIOLETTE_New-LcyE_Forward

tatggcaattaagtcctcagcgagctctaagttggtgtggcggatt

pVIOLETTE_New-LcyE_Reverse

ggttctaaagaatgtcctagttccctctaaa

New-ofCCD1

pVIOLETTE_New-ofCCD1_Forward

attggtctcggcatccgcggcattcc

pVIOLETTE_New-ofCCD1_Reverse

tttgaagctatgcagcgccaccatggcgctgttcgcgct

pADH1-HygR-tVPS13

pFLEUR_hygR_Forward

ggcaggtaccaatccagctagccaatggctatcctttt

pFLEUR_hygR_Reverse

atgcctcagcgtcgacctcattcgcgctgcggatatttct

pCfB3038

pFLEUR_iPCRpCfB3038_Forward

cgcgaatgaggtcgacgctgaggcattaatcttcccgtgaa

pFLEUR_iPCRpCfB3038_Reverse

ttgaaaccctaggttaatgaaagaaccgaaccgatgccaa

Ptef1-Z3ev-tCYC1

pFLEUR_Z3eV_Forward

gttctttcattaacctagggtttcaatacgttatataagttataatgttacat

pFLEUR_Z3eV_Reverse

attggctagctggattggtacctgccgattcatagct

pCUP1-dbr1-tGYP7

pFLEUR_Dbr1_Forward

cgtgtcagaaccatggtgctggaatgggtgaga

pFLEUR_Dbr1_Reverse

atagccattggctagctgttaaattctagttagaaaaagacatttttgct

pZ3eV-Lis-tADH1

pFLEUR_Lis_Forward

tgaatcggcaggtaccgccgtcaccacct

pFLEUR_Lis_Reverse

cattccagcaccatggttctgacacgacaatacatgc

pFLE

pFLEUR_pFle_Forward

gaatttaacagctagccaatggctatccttttgttgtt

pFLEUR_pFle_Reverse

tggtgacggcggtacctgccgattcatagcttcaaa

pAM4951

pCONCOMBRE_pAM4951_Forward

gcggccgcagaaggccatcctgacggatg

pCONCOMBRE_pAM4951_Reverse

gaattcaaaaatgtcgcacaatgtgcgcc

LOX

pCONCOMBRE_LOX_Forward

cacattgtgcgacatttttgaattcgaattctcccttttgttatcaataaaaaag

pCONCOMBRE_LOX_Reverse

ggtgtcatgcatatgcatgtcagaaatattctg

HPL

pCONCOMBRE_HPL_Forward

aatgaaagatctagatcttcccaggaaaaattaaaggg

pCONCOMBRE_HPL_Reverse

cgtcaggatggccttctgcggccgcgcggccgctgtcagaaatattc

LacI

pCONCOMBRE_LacI_Forward

ttctgacatgcatatgcatgacaccatcgaatg

pCONCOMBRE_LacI_Reverse

cctgggaagatctagatctttcattaaagaaagttaaaatgc



Set of primers used for cloning verification


Construction name

Primer pairs name

5’ to 3’ sequence

pFLE

pFLEUR_Z3eV_Forward

gttctttcattaacctagggtttcaatacgttatataagttataatgttacat

pFLEUR_Z3eV_Reverse

attggctagctggattggtacctgccgattcatagct

pFLEUR_hygR_Forward

ggcaggtaccaatccagctagccaatggctatcctttt

pFLEUR_Screen_Reverse

aacttcctaagcatcgcaag

pFLEUR

pFLEUR_Lis_Forward

tgaatcggcaggtaccgccgtcaccacct

pFLEUR_Lis_Reverse

cattccagcaccatggttctgacacgacaatacatgc

pFRAMB

pFRAMBOISEfused_Ptet_Forward

cgcatggaatcgtacgtctctatcactgatagggagtaaactcgaca

pFRAMBOISEfused_Ptet_Reverse

ccactgggtcgactttgtttgtttatgtgtgtttattcgaaactaagttct

pFRAMBOISEfused_neoR_Forward

ataaacaaacaaagtcgacccagtggtctttcagat

pFRAMBOISEfused_neoR_Reverse

tagtcctgcaggttgtggatgatttataatgttacatgcgtacac

pFRAMBOISE-fused

pFRAMBOISEfused_rtTA_Forward

caaactgtgagctagctcacatatgaaagtatatacccgcttttgtaca

pFRAMBOISEfused_rtTA_Reverse

agaccactgggtcgacgctgttctgtccgt

pFRAMBOISEfused_CrtY-phCCD1_Forward

ataaacaaacaaacctaggtgctaaaaaatgcaaccacatt

pFRAMBOISEfused_CrtY-phCCD1_Reverse

tcatatgtgagctagctcacagtttggcttgttcttgaatttgtt

pFRAMBOISE-notfused

pFRAMBOISEnotfused_phCCD1_Forward

agataaggaaaggtcagcaaattctggtg

pFRAMBOISEnotfused_phCCD1_Reverse

tgaagaaaaacgcgctgcggatatttctaag

pFRAMBOISEnotfused_CrtY_Forward

aacaaacaaaatgcaacctcactacgac

pFRAMBOISEnotfused_CrtY_Reverse

ttgctgacctttccttatctttgcattgaac

pVIOLET

pVIOLETTE_Pseq1_Forward

tttatcaaattttgcgttcgag

pVIOLETTE_Screen_Reverse

agggtacaccaaattccacc

pVIOLETTE

pVIOLETTE_Pseq1_Forward

tttatcaaattttgcgttcgag

pVIOLETTE_nsrR_Reverse

ttaatgcctcagcactagtcctgcagagcgccattata

pCONCOMBRE

pCONCOMBRE_LOX_Forward

cacattgtgcgacatttttgaattcgaattctcccttttgttatcaataaaaaag

pCONCOMBRE_LOX_Reverse

ggtgtcatgcatatgcatgtcagaaatattctg

pCONCOMBRE_HPL_Forward

aatgaaagatctagatcttcccaggaaaaattaaaggg

pCONCOMBRE_HPL_Reverse

cgtcaggatggccttctgcggccgcgcggccgctgtcagaaatattc

pCONCOMBRE_LacI_Forward

ttctgacatgcatatgcatgacaccatcgaatg

pCONCOMBRE_LacI_Reverse

cctgggaagatctagatctttcattaaagaaagttaaaatgc



Set of primers used for plasmid linearization


Construction name

Primer pairs name

5’ to 3’ sequence

PpFRAMBOISE-fused
pFRAMBOISE-notfused

pFRAMBOISE_pCfB3032_Forward

acaggcaatactctgcag

pFRAMBOISE_pCfB3032_Reverse

tctctagaaagtataggaacttcac

pVIOLETTE

pVIOLETTE_pCfB3040_Forward

cgcccttattcgactctatag

pVIOLETTE_pCfB3040_Reverse

cgtacctggatggtcatttc

pFLEUR

pFLEUR_pCfB3038_Forward

agaaagtataggaacttctgaag

pFLEUR_pCfB3038_Reverse

catacagcgtgacaataatg



Set of primers used for recombination verification


Construction name

Primer pairs name

5’ to 3’ sequence

pVIOLETTE

897_EasyClone_Forward

gaactgacgtcgaaggctct

pVIOLETTE_iPCRpCfB3040_Reverse

tgtaacattataatggcgctctgcaggactagtgctga

pVIOLETTE_iPCRpCfB3040_Forward

gagctcgctgaggacttaattgccata

898_EasyClone_Reverse

cgtacctggatggtcatttc

pFRAMBOISE-fused
pFRAMBOISE-notfused

903_EasyClone_Forward

tgacgaatcgttaggcacag

pFRAMBOISEfused_iPCRpFRAMB_Reverse

tttagcacctaggtttgtttgtttatgtgtgtttattcgaaactaagttct

pFRAMBOISEfused_iPCRpFRAMB_Forward

acagaacagcgtcgacccagtggtctttcagat

904_EasyClone_Reverse

ccgtgcaataccaaaatcg

pFLEUR

891_EasyClone_Forward

ctggcaagagaaccaccaat

pFLEUR_pFle_Reverse

tggtgacggcggtacctgccgattcatagcttcaaa

pFLEUR_pFle_Forward

gaatttaacagctagccaatggctatccttttgttgtt

892_EasyClone_Reverse

ggacgacaactacggaggat

pCONCOMBRE

pCONCOMBRE_LOX_Forward

cacattgtgcgacatttttgaattcgaattctcccttttgttatcaataaaaaag

pCONCOMBRE_LOX_Reverse

ggtgtcatgcatatgcatgtcagaaatattctg

pCONCOMBRE_HPL_Forward

aatgaaagatctagatcttcccaggaaaaattaaaggg

pCONCOMBRE_HPL_Reverse

cgtcaggatggccttctgcggccgcgcggccgctgtcagaaatattc

pCONCOMBRE_LacI_Forward

ttctgacatgcatatgcatgacaccatcgaatg

pCONCOMBRE_LacI_Reverse

cctgggaagatctagatctttcattaaagaaagttaaaatgc

pCONCOMBRE_RA_Forward

aattgcagttcgcgcttagc

pCONCOMBRE_RA_Reverse

ctgctcccggcatcc

pCONCOMBRE_LA_Forward

ttgatcttttctacggggtctg

pCONCOMBRE_LA_Reverse

ttccaatgccttctccaagg

pCONCOMBRE_specR_Forward

agcatcgcgcgcgcgcgc

pCONCOMBRE_specR_Reverse

ttgagttaagccgcgccgc

DNA bricks sequences



S. cerevisiae gblocks


  • pFRAMBOISE-fused:

    • CrtY-Linker-phCCD1:

      aaaaaatgcaaccacattatgatttaattctcgtgggggctggactagcaaatggtcttatcgccttgcgtcttcaacaacagcaacctgatatgagaatattgcttatcgacgctgcaccccaggctggtggaaatcatacgtggtcatttcaccacgatgatttgactgagtctcaacatagatggatagcgccgctggtggttcatcactggccagactaccaggtccgctttcccacacgccgtagaaagttaaactccggctacttctgtataacttctcaaaggttcgctgaggttttattgcgacaatttggtccacacttgtggatggataccgctgtcgcagaagttaatgctgaatctgttagattgaaaaagggtcaagttatcggtgctagggctgtgattgacgggcggggttatgcggcaaactccgcactcagcgtaggattccaagcttttattggccaggaatggcgattgtcacgaccgcatggtttatcctctccaattatcatggatgcaacggtcgatcagcaaaatggttacaggttcgtgtacagtttgccactctcgccaaccagattgttaatagaagacacacactatatcgataatgcaacattagatcctgaaagggctaggcaaaatatatgcgactacgccgcgcaacagggttggcaacttcagactcttctgagagaagaacaaggtgccttaccgattaccctatctggcaacgctgacgcattctggcaacaaagaccactagcttgtagtggattaagagccggtttgttccatcctaccacaggctattcattgccactggcggttgccgtagccgacagactgagcgcacttgatgtctttacttcggcctcaattcatcaagccattactcattttgcaagagagagatggcagcagcaaaggtttttcagaatgttgaacaggatgttgtttttagctgggcccgccgatttaagatggagagtaatgcaacgtttttatggtttacctgaagatttaattgctagattttatgctggaaaactaacactgacagatcgtctaagaatactaagtggcaaacctcctgttccagtattagcagcattgcaagctattatgacgactcatcgtggtggcggaggcagcggcggaggtggcagcggcggtggcggaagcggtggaggcggtagcatggggagaaaggaaagtgatgatggagtggagagaattgaaggaggagttgttgtagtaaatccaaaaccaaagaagggaattactgctaaggccatagacttattggaaaaagtaatcataaaactaatgcatgattcatcaaaaccacttcactatctttctggcaattttgcacctactgatgaaactccccctcttaatgaccttcctatcaaaggccatcttcctgagtgccttaatggtgaatttgttagagttggtccgaatccaaagtttgctccagttgctggataccactggtttgatggagatggcatgattcacggcttgcgcattaaagatggaaaagcaacgtatgtctcacgttatgtgaggacatcgcgtctaaagcaagaagagttctttgaaggagctaagtttatgaagattggagatcttaaggggctgtttggtctattcacagtttatatgcaaatgctcagagcaaagttgaaaattttggacacctcctatggaaatggcacagctaatacagctctagtatatcatcatgggaagcttttggcactttctgaagctgataaaccttatgcacttaaagttctggaggatggagatcttcaaacacttggcatgctggattatgacaaaagattgttgcattctttcactgcccatccaaaggttgaccctgtaacaggtgagatgtttacctttggctacgctcacgagccgccttacatcacttatagggttatatcaaaagatggaattatgcaggatccggtcccaataacaataccagaggctatcatgatgcatgattttgctattactgaaaattatgcaattatgatggatctccctttgtgtttccggccaaaggaaatggtgaaaaataaccagctggcttttaccttcgatactactaaaaaggctcgatttggagtccttccacgatatgcaaaaagtgaggccctaatcaggtggttcgagcttcctaattgcttcatatttcacaacgccaatgcttgggaggagggagatgaagtcgtgttgattacctgccgccttccgcatccagacctagacatggtcaatggagaagttaaagaaaatcttgagaatttctccaatgagctgtatgaaatgagatttaacatgaagagtggtgcagcttcacagaagaaactttcagagtcttctgttgattttcctcggatcaatgagaactacactggaaggaagcagcgttatgtatatggaaccactttgaacagcattgccaaggtaactggaatcatcaaatttgatttgcatgctgaaccagagactgggaaaaaacagcttgaagttggaggaaatgttcaaggcatctttgatctaggacctggcagatttggttcggaggctgtatttgttccctctcagcctggtacagaatgtgaagaggatgatggttacttgatattcttcgtacatgatgagaacactggaaagtcagctgtgaatgtgattgatgcaaaaacaatgtcagcagagccagtggcagttgttgaactacccaaaagagttccttatggattccatgccttctttgtcacagaggaacaaattcaagaacaagccaaactgtga


    • Pcyc-neoR-Tcyc:

      cagatccgccaggcgtgtatatatagcgtggatggccaggcaactttagtgctgacacatacaggcatatatatatgtgtgcgacgacacatgatcatatggcatgcatgtgctctgtatgtatataaaactcttgttttcttcttttctctaaatattctttccttatacattaggacctttgcagcataaattactatacttctatagacacacaaacacaaatacacacactaaattaataatgattgaacaagatggattgcacgcaggttctccggccgcttgggtggagaggctattcggctatgactgggcacaacagacaatcggctgctctgatgccgccgtgttccggctgtcagcgcaggggcgcccggttctttttgtcaagaccgacctgtccggtgccctgaatgaactgcaagacgaggcagcgcggctatcgtggctggccacgacgggcgttccttgcgcagctgtgctcgacgttgtcactgaagcgggaagggactggctgctattgggcgaagtgccggggcaggatctcctgtcatctcaccttgctcctgccgagaaagtatccatcatggctgatgcaatgcggcggctgcatacgcttgatccggctacctgcccattcgaccaccaagcgaaacatcgcatcgagcgagcacgtactcggatggaagccggtcttgtcgatcaggatgatctggacgaagagcatcaggggctcgcgccagccgaactgttcgccaggctcaaggcgagcatgcccgacggcgaggatctcgtcgtgacccacggcgatgcctgcttgccgaatatcatggtggaaaatggccgcttttctggattcatcgactgtggccggctgggtgtggcggaccgctatcaggacatagcgttggctacccgtgatattgctgaagagcttggcggcgaatgggctgaccgcttcctcgtgctttacggtatcgccgctcccgattcgcagcgcatcgccttctatcgccttcttgacgagttcttctgaacaggccccttttcctttgtcgatatcatgtaattagttatgtcacgcttacattcacgccctcctcccacatccgctctaaccgaaaaggaaggagttagacaacctgaagtctaggtccctatttattttttttaatagttatgttagtattaagaacgttatttatatttcaaatttttcttttttttctgtacaaacgcgtgtacgcatgtaacattata


    • Tvps13-Tgyp7-rtTA-Phxt7:

      tcacatatgaaagtatatacccgcttttgtacactatgtagctataattcaatcgtattattgtagctccgcacgaccatgccttagaaatatccgcagcgcgtttttcttcagtttttccgtttcgggcgggttacggatgtattgatgattatgtggcataaacgaattatatctccggtattcaatatgtaaagttccgtttctatttaccattacccggggagcatgtcaaggtcaaaatcgtcaagagcgtcagcaggcagcatatcaaggtcaaagtcgtcaagggcatcggctgggagcatgtctaagtcaaaatcgtcaagggcgtcggtcggcccgccgctttcgcactttagctgtttctccaggccacatatgattagttccaggccgaaaaggaaggcaggttcggctccctgccggtcgaacagctcaattgcttgtctcagaagtgggggcatagaatcggtggtaggtgtctctctttcctcttttgctacttgatgctcctgttcctccaatacgcagcccagtgtaaagtggcccacggcggacagagcgtacagtgcgttctccagggagaagccttgctgacacaggaacgcgagctgattttccagggtttcgtactgtttctctgttgggcgggtgccgagatgcactttagccccgtcgcgatgtgagaggagagcacagcggtatgacttggcgttgttccgcagaaagtcttgccatgactcgccttccagggggcagaagtgggtatgatgcctgtccagcatctcgattggcagggcatcgagcagggcccgcttgttcttcacgtgccagtacagggtaggctgctcaactcccagcttttgagcgagtttccttgtcgtcaggccttcgataccgacaccattgagtaattccagagcaccgtttatgacgttgctcttgtccagtctcgacattttttgattaaaattaaaaaaactttttgtttttgtgtttattctttgttcttagaaaagacaagttgagcttgtttgttcttgatgttttattattttacaatagctgcaaatgaagaatagattcgaacattgtgaagtattggcatatatcgtctctatttatactttttttttttcagttctagtatattttgtattttcctccttttcattctttcagttgccaataagttacaggggatctcgaaagatggtggggatttttccttgaaagacgactttttgccatctaatttttccttgttgcctctgaaaattatccagcagaagcaaatgtaaaagatgaacctcagaagaacacgcaggggcccgaaattgttcctacgagaagtagtgggtcataaaaagtttattccctggaaaaaaaattttgcgttgcctttctggagaattttttcgaattagcgtgctgccactgcatgcatttctgagaagtgtgggcattcttccaccagttgttcctcctaaaaaaaaaaaagatttcctaccccgcacaaattcctgcatacccctcatttccacgg


  • pFRAMBOISE-notfused:

    • CrtY-Tadh1-Ptef1-fyn:

      atgcaacctcactacgacttgatcttagttggtgctggtttggctaacggtttgatcgctcttagattgcagcaacaacaaccagacatgagaattttgttgattgatgccgctccacaagccggtggtaaccatacctggtctttccaccacgatgacttgactgaatctcaacacagatggattgccccattggtcgttcaccactggccagattaccaagtcagattcccaaccagaagaagaaagttgaactccggttacttctgtatcacctcccaacgttttgctgaagtcctgttaagacaattcggtccacatttgtggatggacacagctgtcgctgaagttaacgctgaatccgttcgtttgaagaagggtcaagttattggtgctagagctgtcattgacggaagaggttatgctgctaactctgctctttccgtcggtttccaagctttcatcggtcaagaatggagattgtcccgtccacacggtttgtcttctccaatcatcatggatgctaccgttgaccaacaaaacggttacagattcgtctactccttgccattgagtcctaccagattattaattgaagacacccattacatcgacaacgccactttggacccagaacgtgccagacaaaacatctgtgactacgctgctcaacaaggttggcaattgcaaactttgttgagagaagaacaaggtgccttgccaattactttgtctggtaatgctgatgctttctggcaacaaagaccactggcttgttcaggtttgagagctggtttattccatccaactactggttactctttgccattggctgttgctgttgccgatagactatccgcattggatgttttcacttctgcttctattcaccaagctattacacacttcgctagagaaagatggcaacaacaaagattcttcagaatgttgaacagaatgttgtttctagctggtccagctgacttgagatggagagttatgcaaagattctacggtctaccagaagatttgatcgctagattttatgccggtaaattaaccttgaccgatcgtttgagaattttgtctggtaagccgccagttccagtcttagctgctttgcaagctatcatgactactcaccgttaagcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagcatgaggtcgctcttattgaccacacctctaccggcatgcatagcttcaaaatgtttctactccttttttactcttccagattttctcggactccgcgcatcgccgtaccacttcaaaacacccaagcacagcatactaaatttcccctctttcttcctctagggtgtcgttaattacccgtactaaaggtttggaaaagaaaaaagagaccgcctcgtttctttttcttcgtcgaaaaaggcaataaaaatttttatcacgtttctttttcttgaaaatttttttttgatttttttctctttcgatgacctcccattgatatttaagttaataaacggtcttcaatttctcaagtttcagtttcatttttcttgttctattacaactttttttacttcttgctcattagaaagaaagcatagcaatctaatctaagatgggttgtgttcaatgcaaagataaggaa


    • Linker-phCCD1-Tvps13:

      aggtcagcaaattctggtgccggtgctggcgcgggagctattttgagtagaatggggagaaaggaaagtgatgacggagtggagagaattgaaggtggagttgttgtagtgaacccaaaaccaaagaagggtattactgctaaggccatagacttattggaaaaagtaatcataaaactaatgcatgattcatcaaaaccattgcactatctttctggcaattttgcgcctaccgatgaaactccccctcttaacgacttacctataaaaggccatcttccagagtgccttaatggtgaatttgttagagttggtccgaatccaaagtttgcgccagttgctggttaccactggtttgatggtgatggcatgattcacggcttgcgcattaaagatggaaaagcaacgtacgtctcacgttacgtgaggacatcgcgtctaaagcaagaagagttctttgaaggagctaagtttatgaagattggtgatctcaaggggctgtttggtttattcacggtttatatgcaaatgctcagagcattgttgaaaattttggacacctcctacggtaatggcacagccaatacagctctagtatatcaccatgggaagttattggcattatccgaagctgataaaccttatgcacttaaagtcctggaggatggtgatttacaaacattaggcatgctggattatgacaaaagattgttgcattctttcactgcccatccaaaggttgaccctgtaacaggtgagatgtttacctttggctacgctcacgaaccgccttacatcacgtatagggtcatatcaaaagacggtattatgcaggatccggtcccaataacaataccagaggcgatcatgatgcatgattttgcgattacggaaaattacgcaattatgatggatctccccttgtgtttccggccaaaggaaatggtgaaaaacaaccaattagcttttaccttcgatactactaaaaaggctagatttggagtcttaccacgatatgcaaaaagtgaggccctaatcaggtggttcgagcttcctaattgcttcatatttcacaacgccaatgcttgggaagaaggagatgaagtcgtgttgattacctgtagattaccgcatccagacctagacatggtcaatggggaagttaaagaaaacttagagaatttctccaatgagctgtatgaaatgagatttaacatgaagagtggtgcagctagtcagaagaaattgtctgaatcttctgttgattttcctcgaatcaatgaaaactacactggaaggaaacagcgttatgtatatggaaccactttgaacagcattgccaaggtaactggtatcatcaaatttgatttgcatgctgaaccagagactgggaaaaaacaattagaagttggtggaaatgttcaaggtatctttgacctaggacctggcagatttggttcggaagctgtatttgttccctctcagcctggtacagaatgtgaagaggatgacggttacttgatattcttcgtacatgatgaaaacactggtaagtcagctgtgaatgtgattgatgcaaaaacaatgagcgcagaaccagtggcagttgttgaattacccaaaagagttccctatggtttccatgccttcttcgtcacagaggaacaaattcaagaacaagccaaactgtaatcacatatgaaagtatatacccgcttttgtacactatgtagctataattcaatcgtattattgtagctccgcacgaccatgccttagaaatatccgcagcgcg


  • pVIOLETTE:

    • Linker-ofCCD1m-Tvps13:

      ggcggtggcggaagcggtggaggcggtagcatgagcgataaaattattcatctgaccgacgatagctttgacaccgatgtgctgaaagcggatggtgcgattctggtggatttttgggcggaatggtgcggcccgtgcaagatgattgcgccgatactggatgagattgcggatgaatatcagggcaaactgaccgtggcgaaactgaacattgatcagaacccgggcaccgcgccgaaatatggcatccgcggcattccgaccctgctactgtttaaaaacggcgaagtggcggcgaccaaggtgggcgcgctgagcaaaggccagctgaaagaatttctggatgcgaatctggcgggccgcaaagaaagcgacgatggcgtggaacgcattgaaggcggcgtcgtggtagtgaacccgaagccgaagaaaggcattaccgcgagcaaggcgattgattggctggaatggctgttcgtgaagatgatgcatgatagcaaacagccgctgcattatctgagcggcaactttgcgccggtggatgaaaccccgccgctgaaggatctgccggtgaccggccatctgccggaatgcctgaacggcgaatttgtgcgcgtgggcccgaatccgaagtttgcgagcattgcgggctatcattggtttgatggagatggcatgattcatgggatgcgcattaaggatggtaaagcgacctatgtgagccgctatgtgcagaccagccgcctcaaacaggaggagttctttggccgcgcgatgtttatgaagattggcgatctgaagggcatgttcggcctgtttaccgtgtatatgcagatgctgcgcgcgaaactgaaggtgctggatgcgagctatggcattggcaccgcgaacaccgcgctggtgtatcatcacggcaaactcctggcgctaagcgaagcggataaaccgtatgcgattaaagtgctggaagatggcgatcttcagaccattggcctgctagattatgataaacgcctggcgcatagctttaccgcgcatccgaaggtggatccgtttaccggcgaaatgttcacctttggctatagccataccccgccgtatgtgacctatcgcgtgattagcaaagatggcgcgatgaacgatccggtgccgattaccgtgagcggcccgattatgatgcatgatttcgcgattaccgaaaactatgcgatttttatggatctgccgctgtattttaaaccgaaggagatggtgaaagataagaagtttattttcagctttgatgcgacccagaaggcgcgctttggcattctgccgcgctatgcgaaaaacgaaagcctgattaaatggtttgagctgccgaactgctttattttccataacgcgaacgcgtgggaagaaggcgatgaagtggtgctgattacctgccgcctggaaaacccggatctggatatggtgaacagcaccgtgaaagaacgcctggataactttaaaaacgaactgtatgaaatgcgctttaacctacagaacggcctggcgagccagaagaagctgagcgtgagcgcggtggattttccgcgcgtgaatgaaagctataccacccgcaaacagcgctatgtgtatggcaccaccctggataacattgcgaaagtgaccggcattattaaatttgatctgcatgcggaaccggaaaccggcaaagaaaaactggaactgggcggcaacgtgaaaggcatttttgatctgggtccgggccgctttggcagcgaagcggtgtttgtgccgcgccatccgggcattaccagcgaagaggatgatggctatctgattttttttgtgcatgatgagaacaccggcaaaagcgcggtgaacgtgattgatgcgaaaaccatgagcccggatccggtggcggtggtggaactgccgaaacgcgtgccgtatggctttcatgcgttttttgtgaccgaagatcagctgcaagaacaggcgaaagtgtagtcacatatgaaagtatatacccgcttttgtacactatgtagctataattcaatcgtattattgtagctccgcacgaccatgccttagaaatatccgcagcgcg


    • Pgal-LcyE-Tvps13:

      cggattagaagccgccgagcgggcgacagccctccgacggaagactctcctccgtgcgtcctcgtcttcaccggtcgcgttcctgaaacgcagatgtgcctcgcgccgcactgctccgaacaataaagattctacaatactagcttttatggttatgaagaggaaaaattggcagtaacctggccccacaaaccttcaaattaacgaatcaaattaacaaccataggatgataatgcgattagttttttagccttatttctggggtaattaatcagcgaagcgatgatttttgatctattaacagatatataaatggaaaagctgcataaccactttaactaatactttcaacattttcagtttgtattacttcttattcaaatgtcataaaagtatcaacaaaaaattgttaatatacctctatactttaacgtcaaggagaaaaaatgaaatgtagtgcaaagtcagatagatgtgttgttgataagcaaggtatttcggtggctgatgaggaagactatgtaaaagctgggggttctgaattgttttttgttcaaatgcaaagaaccaaaagtatggagtcacaatcaaagttaagtgaaaaattggcccagataccaattggcaattgcatactggacttagttgttataggttgtggtccggctggactggcactggctgctgaatcagctaaattgggcttgaatgttggtcttattgggccagatcttcctttcacaaataattacggagtttggcaggacgaatttatcggcctgggcctagaaggttgtatcgaacattcctggaaggacacccttgtttatttagatgatgcggatcccattcgcattggtcgggcttatggtcgtgttcacagagatttattacatgaagaattattgaggagatgtgtggaatccggcgtatcttacttgagttctaaagtcgagcgaataactgaagcaccaaacggttattctttgatagaatgcgaaggtaacataactattccatgtcgtttggcaacagtagcctctggtgctgcctcaggtaaattcctcgaatatgaattagggggccccagagtgtgcgttcaaactgcttacggaattgaggtcgaggtggagaataatccgtacgatccagatttaatggtttttatggattatagggacttcagcaaacacaaaccagaaagtttagaagcgaaatatccaacgtttctttacgtcatggcaatgtcgcctacaaagatctttttcgaagaaacatgcttagcctctagggaggcaatgccgttcaaccttctgaagtctaaactcatgtccagactaaaagctatgggtattagaatcacaagaacttacgaagaagagtggtcgtacatccctgtaggaggtagtcttccaaacaccgaacagaagaacctagcttttggcgcagcggcgtctatggtgcatccagcaactggatatagcgtcgtgagatcattgtccgaagctcctaattatgccgccgttatcgctaagatattacgtcaagaccaatcaaaagagatgatctccttaggaaaatatactaatattagcaagcaagcttgggaaacattgtggcctttagagagaaaacgacagcgcgcattttttttgttcgggctaagccacatagtattaatggatttagagggaactaggacattctttagaaccttttttaggctacctaagtggatgtggtggggttttctcggatcgtccttgtcatctaccgatctgattattttcgcattgtatatgttcgtcattgctcctcattctttaagaatggaacttgtccgtcatttgttaagcgacccaacgggcgccacgatggtaaaagcctacttgactattcccaagggtggcggaggcagcggcggaggtggcagc


    • Ptef1-nsrR-Tcyc1:

      catagcttcaaaatgtttctactccttttttactcttccagattttctcggactccgcgcatcgccgtaccacttcaaaacacccaagcacagcatactaaatttcccctctttcttcctctagggtgtcgttaattacccgtactaaaggtttggaaaagaaaaaagagaccgcctcgtttctttttcttcgtcgaaaaaggcaataaaaatttttatcacgtttctttttcttgaaaattttttttttgatttttttctctttcgatgacctcccattgatatttaagttaataaacggtcttcaatttctcaagtttcagtttcatttttcttgttctattacaactttttttacttcttgctcattagaaagaaagcatagcaatctaatctaagatgggtaccactcttgacgacacggcttaccggtaccgcaccagtgtcccgggggacgccgaggccatcgaggcactggatgggtccttcaccaccgacaccgtcttccgcgtcaccgccaccggggacggcttcaccctgcgggaggtgccggtggacccgcccctgaccaaggtgttccccgacgacgaatcggacgacgaatcggacgacggggaggacggcgacccggactcccggacgttcgtcgcgtacggggacgacggcgacctggcgggcttcgtggtcgtctcgtactccggctggaaccgccggctgaccgtcgaggacatcgaggtcgcaccggagcaccggggtcacggggtcggacgcgcgttgatggggctcgcgacggagttcgcacgcgagcggggtgccggtcacctctggctggaggtcaccaacgtcaacgcaccggcgatccacgcgtaccgacggatggggttcaccctctgcggcctggacaccgccctgtacgacggcaccgcctcggacggcgagcaggcgctctacatgagcatgccctgcccctaaacaggccccttttcctttgtcgatatcatgtaattagttatgtcacgcttacattcacgccctcctcccacatccgctctaaccgaaaaggaaggagttagacaacctgaagtctaggtccctatttattttttttaatagttatgttagtattaagaacgttatttatatttcaaatttttcttttttttctgtacaaacgcgtgtacgcatgtaacattata


    • Final Round:

      tttagagggaactaggacattctttagaaccttttttaggctacctaagtggatgtggtggggttttctcggatcgtccttgtcatctaccgatctgattattttcgcattgtatatgttcgtcattgctcctcattctttaagaatggaacttgtccgtcatttgttaagcgacccaacgggcgccacgatggtaaaagcctacttgactattcccaagggtggcggaggcagcggcggaggtggcagcggcggtggcggaagcggtggaggcggtagcatgagcgataaaattattcatctgaccgacgatagctttgacaccgatgtgctgaaagcggatggtgcgattctggtggatttttgggcggaatggtgcggcccgtgcaagatgattgcgccgatactggatgagattgcggatgaatatcagggcaaactgaccgtggcgaaactgaacattgatcagaacccgggcaccgcgccgaaatatggcat


  • pFLEUR:

    • pADH1-HygR-tVPS13:

      atccttttgttgtttccgggtgtacaatatggacttcctcttttctggcaaccaaacccatacatcgggattcctataataccttcgttggtctccctaacatgtaggtggcggaggggagatatacaatagaacagataccagacaagacataatgggctaaacaagactacaccaattacactgcctcattgatggtggtacataacgaactaatactgtagccctagacttgatagccatcatcatatcgaagtttcactaccctttttccatttgccatctattgaagtaataataggcgcatgcaacttcttttctttttttttcttttctctctcccccgttgttgtctcaccatatccgcaatgacaaaaaaatgatggaagacactaaaggaaaaaattaacgacaaagacagcaccaacagatgtcgttgttccagagctgatgaggggtatctcgaagcacacgaaactttttccttccttcattcacgcacactactctctaatgagcaacggtatacggccttccttccagttacttgaatttgaaataaaaaaaagtttgctgtcttgctatcaagtataaatagacctgcaattattaatcttttgtttcctcgtcattgttctcgttccctttcttccttgtttctttttctgcacaatatttcaagctataccaagcatacaatcaactatgggtaaaaagcctgaactcaccgcgacgtctgtcgagaagtttctgatcgaaaagttcgacagcgtctccgacctgatgcagctctcggagggcgaagaatctcgtgctttcagcttcgatgtaggagggcgtggatatgtcctgcgggtaaatagctgcgccgatggtttctacaaagatcgttatgtttatcggcactttgcatcggccgcgctcccgattccggaagtgcttgacattggggagttcagcgagagcctgacctattgcatctcccgccgtgcacagggtgtcacgttgcaagacctgcctgaaaccgaactgcccgctgttctgcagccggtcgcggaggcaatggatgcgatcgctgcggccgatcttagccagacgagcgggttcggcccattcggaccgcaaggaatcggtcaatacactacatggcgtgatttcatatgcgcgattgctgatccccatgtgtatcactggcaaactgtgatggacgacaccgtcagtgcgtccgtcgcgcaggctctcgatgagctgatgctttgggccgaggactgccccgaagtccggcacctcgtgcacgcggatttcggctccaacaatgtcctgacggacaatggccgcataacagcggtcattgactggagcgaggcgatgttcggggattcccaatacgaggtcgccaacatcttcttctggaggccgtggttggcttgtatggagcagcagacgcgctacttcgagcggaggcatccggagcttgcaggatcgccgcggctccgggcgtatatgctccgcattggtcttgaccaactctatcagagcttggttgacggcaatttcgatgatgcagcttgggcgcagggtcgatgcgacgcaatcgtccgatccggagccgggactgtcgggcgtacacaaatcgcccgcagaagcgcggccgtctggaccgatggctgtgtagaagtactcgccgatagtggaaaccgacgccccagcactcgtccgagggcaaaggaataatcacatatgaaagtatatacccgcttttgtacactatgtagctataattcaatcgtattattgtagctccgcacgaccatgccttagaaatatccgcagcgcg


    • pCUP1-dbr1-tGYP7:

      ctagttagaaaaagacatttttgctgtcagtcactgtcaagagattcttttgctggcatttcttctagaagcaaaaagagcgatgcgtcttttccgctgaaccgttccagcaaaaaagactaccaacgcaatatggattgtcagaatcatataaaagagaagcaaataactccttgtcttgtatcaattgcattataatatcttcttgttagtgcaatatcatatagaagtcatcaaaaaatggaacagcaacaagaagtgatcactaacaagaaagtaatattgaaagactacgttgtagggtttccaaaggagtccgacatgatccttaaaacatcggaaaccatgacattaaagcttccagcaggttcaaatggtttattagttaagaatttatatttgagctgtgatccttacatgaggtctagaatgactaaaactgaaggcagttatgtcgagtcttttactccgggttcacctttaacaggttatggagtagccaaagttttggaatccggtcatgcaaactttaaaaaaggcgacctaatttggggtttcacaggttgggaagagtacagcattatcaatgctcctgagggtctattcaagattgaacataccgatgtgcctctttcttattatacaggcatacttggtatgccaggcatgactgcttatgttggtttctatgagatatgtactccaaaaaaaggagaatatgtctttgtctcggctgcttctggtgcagttggtcagttagtcgggcagtttgctaagttgtccggatgctatgtcgttggtagtgctggtacgaaggaaaaggttgatttgctgaagaacaaatttggattcgatgaagcttttaattacaaagaagaacaagacctggatgccgccctgaagaggtactttccggaaggtattgatatttacttcgagaacgttggcggaagaatgttggatgcagtattgttgaacatgagactagatggcagaatttcagtttgtggtatgatctcacaatacaacttagaacaaagtgagggagtccgaaatctcttcaccctggtaacgaaacgtgtgaccatgaaagggttcattgtgtttgatcactatcacaagtacccaaaatatttagaaatgattatacccctaatcaaaaatggcacgataaattacatagaagacattgtagaagggcttgagaatgcacccgcggccttgattggtttatattctggaaaaaatgttggaaagcaagttgtggtggttgcgcatgaataatggtaaatagaaacggaactttacatattgaataccggagatataattcgtttatgccacataatcatcaatacatccgtaacccgcccgaaacggaaaaactgaagaaaaa


    • Ptef1-Z3ev-tCYC1:

      catagcttcaaaatgtttctactccttttttactcttccagattttctcggactccgcgcatcgccgtaccacttcaaaacacccaagcacagcatactaaatttcccctctttcttcctctagggtgtcgttaattacccgtactaaaggtttggaaaagaaaaaagagaccgcctcgtttctttttcttcgtcgaaaaaggcaataaaaatttttatcacgtttctttttcttgaaaattttttttttgatttttttctctttcgatgacctcccattgatatttaagttaataaacggtcttcaatttctcaagtttcagtttcatttttcttgttctattacaactttttttacttcttgctcattagaaagaaagcatagcaatctaatctaagatggggacccgcccatatgcttgccctgtcgagtcctgcgatcgccgcttttctcgctcggatgagcttacccgccatatccgcatccataccggtcagaagcccttccagtgtcgaatctgcatgcgtaacttcagtcgtagtgaccaccttaccacccacatccgcacccacacaggcgagaagccttttgcctgtgacatttgtgggaggaagtttgccaggagtgatgaacgcaagaggcataccaaaatccatacaggtggcggaggcacacctgcagctgcgtcaactctagaggacccatctgctggagacatgagagctgccaacctttggccaagcccgctcatgatcaaacgctctaagaagaacagcctggccttgtccctgacggccgaccagatggtcagtgccttgttggatgctgagccccccatactctattccgagtatgatcctaccagacccttcagtgaagcttcgatgatgggcttactgaccaacctggcagacagggagctggttcacatgatcaactgggcgaagagggtgccaggctttgtggatttgaccctccatgatcaggtccaccttctagaatgtgcctggctagagatcctgatgattggtctcgtctggcgctctatggagcacccagtgaagctactgtttgctcctaacttgctcttggacaggaaccagggaaaatgtgtagagggcatggtggagattttcgacatgctgctggctacatcatctcggttccgcatgatgaatctgcagggagaggagtttgtgtgcctcaaatctattattttgcttaattctggagtgtacacatttctgtccagcaccctgaagtctctggaagagaaggaccatatccaccgagtcctggacaagatcacagacactttgatccacctgatggccaaggcaggcctgaccctgcagcagcagcaccagcggctggcccagctcctcctcatcctctcccacatcaggcacatgagtaacaaaggcatggagcatctgtacagcatgaagtgcaagaacgtggtgcccctctatgacctgctgctggagatgctggacgcccaccgcctacatgcgcccactagccgtggaggggcatccgtggaggagacggaccaaagccacttggccactgcgggctctacttcatcggagctccacttagacggcgaggacgtggcgatggcgcatgccgacgcgctagacgatttcgatctggacatgttgggggacggggattccccggggccgggatttaccccccacgactccgccccctacggcgctctggatatggccgacttcgagtttgagcagatgtttaccgatgcccttggaattgacgagtacggtgggtagacaggccccttttcctttgtcgatatcatgtaattagttatgtcacgcttacattcacgccctcctcccacatccgctctaaccgaaaaggaaggagttagacaacctgaagtctaggtccctatttattttttttaatagttatgttagtattaagaacgttatttatatttcaaatttttcttttttttctgtacaaacgcgtgtacgcatgtaacattata


    • pZ3eV-Lis-tADH1:

      ttatattgaattttcaaaaattcttactttttttttggatggacgcaaagaagtttaataatcatattacatggcattaccaccatatacatatccatatctaatcttacttatatgttgtggaaatgtaaagagccccattatcttagcctaaaaaaaccttctctttggaactttcagtaatacgcttaactgctcattgctatattgaagtggcgtgggcgccgtgcgtcctcgtcttcaccggtcgcgttcctgaaacgcagatgtgcctcgcgccgcactgctccgaacaataaagattctacaatactagcttttatggttatgaagaggaaaaattggcagtaacctggccccacaaaccttcaaattaacgaatcaaattaacaaccataggatgataatgcgattagttttttagccttatttctggggtaattaatcagcgaagcgatgatttttgatctattaacagatatataaatggaaaagctgcataaccactttaactaatactttcaacattttcagtttgtattacttcttattcaaatgtcataaaagtatcaacaaaaaattgttaatatacctctatactttaacgtcaaggagaaaaaactataaaaaaatgtcgatcaatatcaacatgcctgcagccgccgtcctccgcccttttcgctgctcacaactacatgtcgatgaaacccgacgctccggaaactaccgcccctcggcttgggattccaactacatccaatctctcaattctcagtataaggaaaagaagtgcttgacaaggctagaagggctgattgagcaagtgaaggaactgaaggggacaaaaatggaggctgttcaacaattggagttgattgatgactcgcagaatctgggattatcatattactttcaagataaaattaaacatatcttgaatttgatatataatgatcacaaatatttttacgatagtgaagctgaaggaatggatttgtattttacagctcttggatttagactctttagacaacatggttttaaagtctcccaagaagtatttgatcgtttcaagaacgagaatggtacgtatttcaagcacgacgatacaaagggattgttgcagctctacgaagcatcattcctagtgcgagaaggcgaagagacactcgaacaagcacgagaatttgccaccaaatccctacaaagaaaacttgatgaggatggtgatggaattgacgccaatatcgaatcatggatacgccactctctggagatcccacttcattggagggctcagaggctagaggcgagatggttcctagatgcttatgcgagaaggcccgacatgaaccccgttatcttcgagcttgctaaactcaacttcaatattgtccaagcaacacaacaagaagaattgaaagctctttcgaggtggtggagtagtttaggcctagctgaaaaactcccatttgtgagggataggcttgtggaaagctacttttgggctattccactctttgagcctcatcaatatggatatcaaagaaaagtggccaccaagatcataaccctaatcacatctttagacgatgtttacgatatctatggcacgttagatgaattgcaactatttacgaacttgtttgaaagatgggataatgcatcaatcggccgacttcctgaatacttgcaattgttctatttcgcaatccacaactttgtttccgaggtggcttacgacattctcaaagaaaagggtttcactagtattgtatatttacagagatcgtgggtggatttgctaaaaggatacctaaaagaggcaaagtggtacaatagtggatacacgccaagcctagaggaatatttcgacaacgcattcatgacaataggggcccctccggtactatcgcaagcttacttcacattaggaagctcgatggagaaaccgatcatcgagagcatgtacgaatatgacaacatacttcgcgtttcgggaatgctcgtgaggcttcccgatgaccttggaacatcatcgttcgagatggagagaggcgacgtgccgaaatcggtccagctatacatgaaggaaacaaatgctacggaggaggaggcggtggagcacgtgaggtttttgaatcgggaggcgtggaagaagatgaacacggcggaggcggccggtgattctccgttagtgagtgacgtggtggcggtggcggcgaatcttggaagggcggcgcagtttatgtatttcgacggagatggtaaccagtctagtttgcagcagtggattgtgagcatgctgttcgagccgtaggcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagcatgaggtcgctcttattgaccacacctctaccggcatg



S. elongatus gblocks


  • pCONCOMBRE:

    • LOX:

      aaatattctgaaatgagctgttgacaattaatcatccggctcgtataatgtgtggaattgtgagcggataacaatttcatacgctcacaattggtaccggtgataccagcatcgtcttgatgcccttggcagcaccctgctaaggaggcaacaagatgtccttggagaagatcgtggatgccattagtggtaaggatgacgggaaaaaggttaaagggacggtggttctgatgaaaaaaaacgtactcgattttactgatattaatgccagtgttctcgatggcgtcctggagttcttgggtcgacgggtgtccctcgagttaatctcgtcggtgcacgttgatcctgcaaatggcctacaaggtaagcgctccaaggcagcgtacctcgaaaactggctgaccaacaagactcccatcgcggcaggagaatcggcgtttcgcgttacctttgactgggacgatgaggagtttggagtgcctggggctttcatcattaagaacttgcactttagcgagttcttcctcaagtcgctgacgctcgaagatgtcccgaaccacgggaaagtccattttgtctgcaatagttgggtctatccagccaacaaatataagtcgccacgcattttttttgctaatcaggcgtacctcccgtccgaaactcccgaaccactgcgcaaatgtcgcgagaacgaactggtgactcttcgcggtgacgggacaggcaaacttgaagagtgggatcgcgtgtatgactatgcctactacaacgaccttggcgatccggacaaaggcaaggagttaagccgccctgtcctcggggggagttccgaatatccctacccgcggcggggccgcacgggtcgtgagcccaccaagagcgatccaaactccgagtcccggattcccctcttgatgtctcttgacatctacgttcctcgcgacgagcgctttggccatattaaactgtcggacttcctgaccttcgcgcttaaatcgatcgtccagctactactgccggagttccaagccctgtttgatagtacgcccaatgagttcgatagttttgaagacgtgctgaagctctatgaaggcgggataaaactcccccaaggcccgctgctcaaagcgatcaccgataatattcctttggaaattctcaaagagttactgcgatcggatggagaaggcctgttcaagtacccaacccctcaagtgattcaagaggataaaacggcttggcggactgacgaggaatttggtcgggagatgctggctggcgtcaacccggtggtcatctcgagactccaggaatttcctccgaagagcaaattggatcccaaaacctacggcaaccagaactcgacgattactcgcgaacaaattgaggataagcttgatggcttgacgatcgatgaagccattaaaaccaataaactgtttattttgaatcatcacgatattctgatgccctatctgcgtcgcattaatacctctacagataccaagacctacgcgtcacgcaccctgttatttctccaagataatggaacattgaaaccattagccatcgaactgagcctaccccatcccgatggtgatcagtttggcgctgtcagtaaagtctacaccccagctgatcagggcgtggaaggcagtatttggcagttggccaaagcctatgctgccgtgaacgattcaggagttcatcagctgatcagccattggttgaatacccacgcggtcatcgaaccctttgtgatcgctactaaccgccagctgtcaacactccacccgatctacaaactgttgcacccgcacttccgagaaacgatgaacattaatgcactcgctcgtcagatcctcatcaacggcggaggcctgctggaactgaccgtcttcccggccaaatacagcatggaaatgtcagctgtagtttataaagattgggtatttccagagcaagcattgccgactgacctgatcaagcgaggggtggcagttgaggactcatcgagtccacatggcattcgtttactgattcaggattatccctatgccgttgatggacttaagatttggagcgcgatcaagtcttgggtgacggagtactgcaactattactacaagagcgatgatgcggtgcaaaaagatactgaattgcaagcttggtggaaggagctgcgggaagagggccatggcgacaagaaggacgaaccatggtggccgaagatgcagacggtacaggaattgattgacagctgtacgattaccatctggattgcctctgcgctgcatgcagcggttaacttcggtcaatacccttacgcgggttatctgcctaatcgacctactcttagccgcaagttcatgccggaacctggctcgcccgcatatgaagagctaaaaacgaatcccgataaggtctttctcgaaacaatcacgccgcagttgcaaaccttgctcggcatcagtcttattgaaattctgtcccgccacagcagcgatacgctctacctagggcagcgcgaaagccccgagtggacaaaagaccaagagccgttgtctgcttttgggaggttcggtaagaaactctcggacatcgaagatcaaatcatgcagatgaatggcgatgaaaaatggaaaaatcgtagtggtcccgtcaaagtgccgtatacgttgctattccccacctcagagggtggtttgaccggcaagggcattccgaatagtgtcagcatctaagacgaacaataaggcctcccaaatcggggggccttttttattgataacaaaa


    • HPL:

      aaatattctgaaatgagctgttgacaattaatcatccggctcgtataatgtgtggaattgtgagcggataacaatttcatacgctcacaattggtaccggtgataccagcatcgtcttgatgcccttggcagcaccctgctaaggaggcaacaagatggctacaccgtcgagtagcagtcctgagctccctttgaaaccgatcccaggtgggtatggctttccgttcctaggtcctatcaaggatcgttacgactacttctattttcagggtcgcgatgaatttttccggagccgcatcaccaagtacaacagcaccgtttttcgggccaacatgcctcccggaccattcatctcatctgactcgcgcgtcgtcgtgctgctggatgcgttgtcgttcccgattttgtttgatacggccaaagttgaaaaacggaatatccttgatggcacttacatgccgtccctcagcttcaccggcaatattcgcacttgcgcctatctcgatcccagcgaaactgagcattcggtgctcaagcgcctgtttctctcgttccttgcttctcgccatgatcgcttcattccgctcttccgctcgtcattatcagaaatgttcgtcaagctagaagacaagttgagtgagaagaagaaaatcgctgacttcaatagcattagcgattccatgagttttgactatgtttttcgactgttaagcgacggtacgcccgattccaaactggcagcggagggcccaggcatgtttgatctctggctggtcttccaactagctcccctcgcgtcgatcggtttgcccaaaatcttcagcgtctttgaagatttggtgattcacacgattcccctgccgttctttcctgtaaaaagtggctatcgcaaactctacgaagccttttactcttctagtgggtcttttttggatgaggcagagaagcaaggcatcgatcgcgaaaaagcctgtcacaacctggtgtttttggcaggctttaacgcctacggcggaatgaaggtcctgttccccaccctccttaaatgggtggggacagcgggagaggatttacatcgtaagctggctgaggaagtgcgtaccacagtgaaagaagagggtggcctgacctttagtgcgctggaaaagatgagtttgctgaagtcggtagtctacgaggctctacgcattgaaccgcccgtcccatttcaatatggaaaagcaaaggaagatattgtgattcagagccacgattcatcctttaagatcaaaaagggggaaacgatctttggctaccagccctttgccaccaaagatccgaagatttttaaagactccgaaaaatttgtgggcgaccgttttgtcggtgaagaaggcgagaagctgctgaaatacgtttattggagcaatgagcgggaaactgtcgagccaacgccggaaaacaagcagtgccccggcaaaaatttggtcgttctgattggtcggatcatggtggtggagttcttcttgcgatatgacaccttcacggttgaggttgcggatctaccacttggacccgcggtgaagttcaaatccctcactcgagcaaccgacatggtctaaagttaaccaaaaaggggggattttatctcccctttaatttttcct

Go to Protocols