Synthetic Biology:Vectors/Parts
This is a list of BioBricks parts for use in construction of modular vectors.
Some have just been designed while others have been constructed and tested. See the associated registry page for details.
Replication origins
- <bbpart>BBa_I50000</bbpart>: F plasmid backbone with BioBricks restriction sites removed.
- <bbpart>BBa_I50001</bbpart>: F plasmid backbone with BioBricks restriction sites removed, reverse orientation.
- <bbpart>BBa_I50010</bbpart>: oriV origin which requires TrfA protein to be functional.
- <bbpart>BBa_I50020</bbpart>: high copy origin of replication from pSB1A3
- <bbpart>BBa_I50030</bbpart>: a pBR322 origin.
- <bbpart>BBa_I50040</bbpart>: a near minimal pSC101 origin.
- <bbpart>BBa_I50041</bbpart>: a near minimal pSC101 origin, reverse orientation.
- <bbpart>BBa_I50050</bbpart>: a R6K gamma origin. See also EZ-Tn5™ <R6Kγori /KAN-2> Sequence general information (Note: this origin will only replicate in a pir+ strain.)
Antibiotic resistance cassettes
- <bbpart>BBa_P1001</bbpart>: cassette providing tetracycline resistance.
- <bbpart>BBa_P1000</bbpart>: cassette providing chloramphenicol resistance.
- <bbpart>BBa_P1002</bbpart>: cassette providing ampicillin resistance.
- <bbpart>BBa_P1003</bbpart>: cassette providing kanamycin resistance.
- <bbpart>BBa_P1004</bbpart>: cassette providing ampicillin resistance in reverse orientation.
- <bbpart>BBa_P1005</bbpart>: cassette providing tetracycline resistance and ampicillin resistance with terminators.
- <bbpart>BBa_P1006</bbpart>: cassette providing chloramphenicol resistance and ampicillin resistance with terminators.
- <bbpart>BBa_P1007</bbpart>: cassette providing kanamycin resistance and ampicillin resistance with terminators.
Terminators
- <bbpart>BBa_B0055</bbpart>: upstream flanking terminator
- <bbpart>BBa_B0054</bbpart>: downstream flanking terminator
- <bbpart>BBa_B0053</bbpart>: bidirectional terminator from E. coli his operon
- <bbpart>BBa_B0052</bbpart>: forward terminator
- <bbpart>BBa_B0062</bbpart>: reverse terminator of BBa_B0052.
Notes
It wasn't clear from the website if these were bi-directional? Not sure that this is very important--BC
- I don't know if the terminators are bidirectional. These terminators are used as flanking terminators in other vectors and are claimed to make the cloning of difficult pieces of DNA (like strong promoters) easier. This is why I was planning on orienting the antibiotic resistance cassette in the opposite direction so that read through from upstream of the multiple cloning site is less of an issue. -- RS
Primer binding sites
- <bbpart>BBa_G00100</bbpart>: VF2
- <bbpart>BBa_G00102</bbpart>: VR
Others
- <bbpart> BBa_I1000</bbpart> (old) or <bbpart>BBa_P1010</bbpart> (preferred): ccd operon in BioBricks format
- <bbpart>BBa_B0042</bbpart>: translational stop sequence which provides a stop codon in all six frames (see Non-functional DNA sequences)
- <bbpart>BBa_B0043</bbpart>: forward Topoisomerase I cloning site (see Topoisomerase I mediated TA cloning)
- <bbpart>BBa_B0044</bbpart>: reverse Topoisomerase I cloning site (see Topoisomerase I mediated TA cloning)
- <bbpart>BBa_G00000</bbpart>: BioBricks prefix
- <bbpart>BBa_G00001</bbpart>: BioBricks suffix
- <bbpart>BBa_B0045</bbpart>: antibiotic cassette insertion site (see Synthetic Biology:Vectors/Modular construction scheme)
- <bbpart>BBa_B0046</bbpart>: replication origin insertion site (see Synthetic Biology:Vectors/Modular construction scheme)
- unique identifier for the vector
Minimal vector scaffold
5' <counter-selectable marker> -- <high copy origin> -- BBa_G00001 (BBsuffix) -- BBa_B0044 (TOPO site) -- BBa_B0042 (translational stop sequence) -- <plasmid barcode> -- BBa_B0054 (terminator) -- BBa_G00102 (VR) -- BBa_B0045 (antibiotic resistance insertion site) -- BBa_B0046 (origin insertion site) -- BBa_G00100 (VF2) -- BBa_B0055 (terminator) -- <plasmid barcode> -- BBa_B0042 (translational stop sequence) -- BBa_B0043 (TOPO site) -- BBa_G00000 (BB prefix) 3'
Ordering information
Part number | Description | Size | BioBrick available | Template available | Want synthesized as an individual part |
BASIC PARTS | |||||
BBa_P1010 | ccd | 675bp | Yes | Yes | No |
BB suffix | 21bp | No | No | No (can make with primers) | |
BBa_B0044 | TOPO site | 13bp | No | No | No (can make with primers) |
BBa_B0042 | translational stop sequence | 12bp | No | No | No (can make with primers) |
plasmid barcode | |||||
BBa_B0054 | terminator | 69bp | No | No | Maybe (can be hard to make terminators with primer extension) |
BBa_G00102 | VR | 20bp | No | No | No (can make with primers) |
BBa_I50000 | F plasmid origin | 4640bp | No | No | No |
BBa_I50001 | F plasmid origin, reverse orientation | 4640bp | No | No | Yes (has 4 BioBricks sites throughout the gene) |
BBa_I50020 | high copy origin (pUC19) | 858bp | No | Yes | Maybe |
BBa_I50040 | low copy origin (pSC101) | 2215bp | No | Yes | No |
BBa_I50041 | low copy origin (pSC101) | 2215bp | No | Yes | Maybe (redundant with BBa_I50030, have template) |
BBa_I50030 | low copy origin (pBR322) | 2016bp | No | Yes | Maybe (redundant with BBa_I50040) |
BBa_P1000 | CmR | 789bp | Yes (Austin) | Yes | No |
BBa_P1001 | TetR | 1279bp | Yes (Austin) | Yes | No |
BBa_P1003 | KanR | 993bp | Maybe (TK) | Yes | Maybe |
BBa_B0053 | His terminator | 72bp | No | No | Yes (previous synthesis attempt via primer extension failed) |
BBa_B0062 | reverse rrnC terminator | 41bp | No | No | Maybe (can be hard to make terminators with primer extension) |
BBa_P1004 | reverse AmpR | 941bp | No | Yes | Maybe |
BBa_G00100 | VF2 | 20bp | No | No | No (can make with primers) |
BBa_B0055 | terminator | 78bp | No | No | Maybe (can be hard to make terminators with primer extension) |
plasmid barcode | |||||
BBa_B0042 | translational stop sequence | 12bp | No | No | No (can make with primers) |
BBa_B0043 | forward TOPO site | 13p | No | No | No (can make with primers) |
BB prefix | 22bp | No | No | No (can make with primers) | |
COMPOSITE PARTS | |||||
BBa_P1005 | tetR and ampR | 1867bp | No | No | Maybe |
BBa_P1006 | cmR and ampR | 2357bp | No | No | Maybe |
BBa_P1007 | kanR and ampR | 2071bp | No | No | Maybe |