Plasmid_Backbone

Part:pSB6A1

Designed by: Stefan Luzi   Group: iGEM06_ETHZ   (2007-10-23)
Revision as of 08:18, 25 October 2007 by Stefan Luzi (Talk | contribs)

No part name specified with partinfo tag.

Plasmid pBR322BB1 (Fig. 1) is based on pBR322 (Fig. 2), a low-copy cloning vector (15-20 copies per cell) that confers ampicillin (ApR) and tetracylin (TcR) resistance and harbours the pMB1 origin of replication.

Fig. 1: Map of the pBR322BB1 plasmid
Fig. 2: Map of the original pBR322 plasmid

In order to make this plasmid compatible with the BioBrick Standard assembly process, the original pBR322 was digested at its EcoRI and BamHI restriction sites and an appropriate multiple cloning site (MCS) was introduced in this position. However, this leads to a loss of the original tetracyclin resistance and only ampicillin resistance is left. Since the original plasmid also contains a PstI restriction site in the bla gene, the GCA codon at this position was changed to GTA by site directed mutagenesis. The term "BB1" in pCK01BB1 refers to "BioBrick version 1".

Purpose

This plasmid was designed for the [http://parts.mit.edu/igem07/index.php/ETHZ ETHZ iGEM 2007 project] and is used for the following BioBricks: Part 4, Part 5new, Part 8, Part 9 and the composites BBa_I739015, BBa_I739016 and BBa_I739017. More information on the plasmid and the used assembly strategy can be found [http://parts.mit.edu/igem07/index.php?title=ETHZ/Biology/Lab here].


Sequence and Features No part name specified with partinfo tag.


[edit]
Categories
//plasmidbackbone/assembly
//plasmidbackbone/copynumber/medium
//plasmidbackbone/operation
Parameters
chassis
copies15-20
mcsBioBrick
originpMB1
resistanceA