Difference between revisions of "Template:TypeIIS/Plasmids/Level1"

Latest revision as of 19:53, 23 June 2020

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).

@@ Line 1: / Line 1: @@
+====Assembling into Level 1 Plasmds====
+pSB1K01-pSB1K04 series of plasmids are based on the pOdd set of Loop Vectors and are built upon the <partinfo>pSB1K3</partinfo> plasmid backbone.
+'''Note:'''
+*Since <partinfo>pSB1K3</partinfo> 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 <partinfo>BBa_J04454</partinfo> have been verified.
 <center>
 [[File:TypeIIS-Level1-J04454-Simple.png]]<br>
@@ Line 14: / Line 22: @@
 <center>[[File:TypeIIS-Level1-TU-Simple.png]]</center>
@@ Line 61: / Line 71: @@
 </html>
+====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
+<center>
+[[File:TypeIIS-Level1-Plasmid-Set.png]]<br>
-'''Note:'''
+'''Simplified diagram of pSB1K0# set with assembled transcriptional units (TUs).'''
-*Since <partinfo>pSB1K3</partinfo> is a high-copy plasmid backbone, these vectors should not be used for Level 3 assemblies.
+</center>
-*Currently, Registry samples of these plasmid backbones have not been fully sequenced. Only their prefix and suffix along with their insert <partinfo>BBa_J04454</partinfo> have been verified.