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_B0043</bbpart>: reverse Topoisomerase I cloning site (see Topoisomerase I mediated TA cloning)
• 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) -- <antibiotic resistance> -- <origin, reverse> -- 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