The main goal of our project is to make a frugal biofoundry capable of producing high quantities of enzymes and other proteins of interest. To do that, our team designed a library of parts in order to test in a high-throughput way the combinations that would generate the best results. For this year, the most important part generated by our team was the pHT43 plasmid (BBa_K4103000), an improvement on (BBa_K1074001) from 2013 - this plasmid acts as a shuttle vector between E. coli and B. subtilis, and allows the efficient production and secretion of our protein-of-interest, with our team focusing efforts to use this vector for the production of Pfu-Sso7d Polymerase (BBa_K4103011) codon optimized for B. Subtilis using the software created by our team.
Part Collection
Overview
Basic Parts
Part Name | Type | Description | Length (bp) |
---|---|---|---|
http://parts.igem.org/Part:BBa_K4103000 | pfu | ||
http://parts.igem.org/Part:BBa_K4103001 | Conjugation origin | Transfer Origin pIP501 | 319 |
http://parts.igem.org/Part:BBa_K4103003 | Resistance marker | SpecR | 989 |
http://parts.igem.org/Part:BBa_K4103004 | Origin of replication | EcoORI | 589 |
http://parts.igem.org/Part:BBa_K4103005 | Origin of transfer | OriTBsub | 378 |
http://parts.igem.org/Part:BBa_K4103007 | Resistance marker | catA9 | 939 |
http://parts.igem.org/Part:BBa_K4103008 | Insulator | HDV-1 (3' Ribozyme Insulator) | 85 |
http://parts.igem.org/Part:BBa_K4103009 | |||
http://parts.igem.org/Part:BBa_K4103016 | OriTBsub | ||
http://parts.igem.org/Part:BBa_K4103017 | Terminator | E. coli terminator | 54 |
http://parts.igem.org/Part:BBa_K4103018 | Promoter | kanR promoter | 101 |
http://parts.igem.org/Part:BBa_K4103019 | Terminator | kanR terminator | 48 |
http://parts.igem.org/Part:BBa_K4103020 | Coding | kanR cds | 816 |
http://parts.igem.org/Part:BBa_K4103021 | Coding | lacI cds | 1083 |
http://parts.igem.org/Part:BBa_K4103022 | |||
http://parts.igem.org/Part:BBa_K4103023 | Operator | Lac operator | 25 |
http://parts.igem.org/Part:BBa_K4103024 | Insulator | RiboJ54 Insulator | 75 |
http://parts.igem.org/Part:BBa_K4103025 | |||
http://parts.igem.org/Part:BBa_K4103026 | Coding | Pfu (DNA polymerase) #4 | 2322 |
http://parts.igem.org/Part:BBa_K4103027 | Coding | Sso-7d #4 | 189 |
http://parts.igem.org/Part:BBa_K4103028 | Linker | TEV linker site #4 | 21 |
http://parts.igem.org/Part:BBa_K4103029 | Coding | fuGFP #4 | 732 |
http://parts.igem.org/Part:BBa_K4103030 | Tag | 10xHisTag #4 | 30 |
http://parts.igem.org/Part:BBa_K4103031 | Tag | R5 Silica binding purification tag #4 | 57 |
http://parts.igem.org/Part:BBa_K4103032 | Coding | Pfu (DNA polymerase) #2 | 2322 |
http://parts.igem.org/Part:BBa_K4103033 | Coding | Sso-7d #2 | 189 |
http://parts.igem.org/Part:BBa_K4103034 | Linker | TEV linker site | 21 |
http://parts.igem.org/Part:BBa_K4103035 | Coding | fuGFP | 732 |
http://parts.igem.org/Part:BBa_K4103036 | Purification Tag | 10xHisTag | 30 |
http://parts.igem.org/Part:BBa_K4103037 | Purification Tag | R5 Silica binding purification tag | 57 |
http://parts.igem.org/Part:BBa_K4103038 | Coding | ccdb | 335 |
http://parts.igem.org/Part:BBa_K4103039 | Homology Arm | lacA 3' homology | 408 |
http://parts.igem.org/Part:BBa_K4103040 | Homology Arm | amyE 5' homology | 521 |
http://parts.igem.org/Part:BBa_K4103041 | Homology Arm | amyE 3' homology | 498 |
http://parts.igem.org/Part:BBa_K4103042 | Homology Arm | thrC 5' homology | 501 |
http://parts.igem.org/Part:BBa_K4103043 | Homology Arm | thrC 3' homology | 429 |
http://parts.igem.org/Part:BBa_K4103044/ | Homology Arm | lacA 5' homology arm | 602 |
http://parts.igem.org/Part:BBa_K4103045 | Secretion Tag | AmyQ | 93 |
Composite Parts
Part Name | Type | Description | Length (bp) |
---|---|---|---|
http://parts.igem.org/Part:BBa_K4103000 | pfu | ||
http://parts.igem.org/Part:BBa_K4103001 | Conjugation origin | Transfer Origin pIP501 | 319 |
http://parts.igem.org/Part:BBa_K4103003 | Resistance marker | SpecR | 989 |
http://parts.igem.org/Part:BBa_K4103004 | Origin of replication | EcoORI | 589 |
http://parts.igem.org/Part:BBa_K4103005 | Origin of transfer | OriTBsub | 378 |
http://parts.igem.org/Part:BBa_K4103007 | Resistance marker | catA9 | 939 |
http://parts.igem.org/Part:BBa_K4103008 | Insulator | HDV-1 (3' Ribozyme Insulator) | 85 |
http://parts.igem.org/Part:BBa_K4103009 | |||
http://parts.igem.org/Part:BBa_K4103016 | OriTBsub | ||
http://parts.igem.org/Part:BBa_K4103017 | Terminator | E. coli terminator | 54 |
http://parts.igem.org/Part:BBa_K4103018 | Promoter | kanR promoter | 101 |
http://parts.igem.org/Part:BBa_K4103019 | Terminator | kanR terminator | 48 |
http://parts.igem.org/Part:BBa_K4103020 | Coding | kanR cds | 816 |
http://parts.igem.org/Part:BBa_K4103021 | Coding | lacI cds | 1083 |
http://parts.igem.org/Part:BBa_K4103022 | |||
http://parts.igem.org/Part:BBa_K4103023 | Operator | Lac operator | 25 |
http://parts.igem.org/Part:BBa_K4103024 | Insulator | RiboJ54 Insulator | 75 |
http://parts.igem.org/Part:BBa_K4103025 | |||
http://parts.igem.org/Part:BBa_K4103026 | Coding | Pfu (DNA polymerase) #4 | 2322 |
http://parts.igem.org/Part:BBa_K4103027 | Coding | Sso-7d #4 | 189 |
http://parts.igem.org/Part:BBa_K4103028 | Linker | TEV linker site #4 | 21 |
http://parts.igem.org/Part:BBa_K4103029 | Coding | fuGFP #4 | 732 |
http://parts.igem.org/Part:BBa_K4103030 | Tag | 10xHisTag #4 | 30 |
http://parts.igem.org/Part:BBa_K4103031 | Tag | R5 Silica binding purification tag #4 | 57 |
http://parts.igem.org/Part:BBa_K4103032 | Coding | Pfu (DNA polymerase) #2 | 2322 |
http://parts.igem.org/Part:BBa_K4103033 | Coding | Sso-7d #2 | 189 |
http://parts.igem.org/Part:BBa_K4103034 | Linker | TEV linker site | 21 |
http://parts.igem.org/Part:BBa_K4103035 | Coding | fuGFP | 732 |
http://parts.igem.org/Part:BBa_K4103036 | Purification Tag | 10xHisTag | 30 |
http://parts.igem.org/Part:BBa_K4103037 | Purification Tag | R5 Silica binding purification tag | 57 |
http://parts.igem.org/Part:BBa_K4103038 | Coding | ccdb | 335 |
http://parts.igem.org/Part:BBa_K4103039 | Homology Arm | lacA 3' homology | 408 |
http://parts.igem.org/Part:BBa_K4103040 | Homology Arm | amyE 5' homology | 521 |
http://parts.igem.org/Part:BBa_K4103041 | Homology Arm | amyE 3' homology | 498 |
http://parts.igem.org/Part:BBa_K4103042 | Homology Arm | thrC 5' homology | 501 |
http://parts.igem.org/Part:BBa_K4103043 | Homology Arm | thrC 3' homology | 429 |
http://parts.igem.org/Part:BBa_K4103044/ | Homology Arm | lacA 5' homology arm | 602 |
http://parts.igem.org/Part:BBa_K4103045 | Secretion Tag | AmyQ | 93 |