Template:TypeIIS/Plasmids

Universal Acceptor Vector (for Level 0)

• pSB1C00 with BBa_J04452

Assembling into Level 1 Plasmds

pSB1K01-pSB1K04 series of plasmids are based on the pOdd set of Loop Vectors and are built upon the pSB1K3 plasmid backbone.

Note:

• Since pSB1K3 is a high-copy plasmid backbone, these vectors should not be used for Level 3 assemblies.
• Currently, Registry samples of these plasmid backbones have not been fully sequenced. Only their prefix and suffix along with their insert BBa_J04454 have been verified.

Simplified diagram of BBa_J04454 and pSB1K0# with BsaI cut site.

When four Level 0 parts are assembled into the plasmid backbone (paired with BBa_J04454) in a one-pot assembly with BsaI...

the resulting transcriptional unit (TU) will be flanked by SapI restriction sites and the specified 3 bp Fusion Sites (NNN).

Level 1 Plasmid Set

Level 1 transcriptional units in this set of plasmid backbones will be flanked by SapI recognition sites and specified 3bp fusion sites, enabling further assembly of up to 4 transcriptional units as a Level 2 Assembly.

Registry Name	Loop Name	Fusion Site 5'	TU	Fusion Site 3'
pSB1K01	pOdd1	ATG	TU 1	GCA
pSB1K02	pOdd2	GCA	TU 2	TAC
pSB1K03	pOdd3	TAC	TU 3	CAG
pSB1K04	pOdd4	CAG	TU 4	GGT

Level 1 -> Level 2 Assembly

Up to 4 Level 1 transcriptional units can be assembled into a Level 2 multi-transcriptional unit. By planning your assemb

Simplified diagram of pSB1K0# set with assembled transcriptional units (TUs).

pEven Loop Vectors (for Level 2)

When four Level 1 parts (TUs) are assembled into a Multi-Transcriptional Unit (MTU) into the following plasmid backbones (with BBa_J04455), the MTU will be flanked by BsaI restriction sites and these 4 bp Fusion Sites.

Registry Name	Loop Name	Fusion Site 5'	MTU	Fusion Site 3'
pSB3C01	pEven1	GGAG	MTU 1	TACT
pSB3C02	pEven2	TACT	MTU 2	AATG
pSB3C03	pEven3	AATG	MTU 3	GCTT
pSB3C04	pEven4	GCTT	MTU 4	CGCT

Note:

• The documented sequence shows an additional BsaI site. This would impact assembly out of (but not into) this set of vectors. A new set of vectors should be created or used in the long term.
• Currently, Registry samples of these plasmid backbones have not been fully sequenced. Only their prefix and suffix along with their insert BBa_J04455 have been verified.