Plasmid_Backbone
pSB8K15FRT

Part:BBa_K864022

Designed by: Erik Lundin   Group: iGEM12_Uppsala_University   (2012-09-24)
Revision as of 22:15, 24 September 2012 by Erlu2699 (Talk | contribs)

Low copy BioBrick temperature sensitive standard vector

pSB8K15(FRT) is a BioBrick standard vector with low copy pSC101ts replication origin (~5 copies per cell) and kanamycin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The pSC101ts is a temperature sensitive pSC101 repelication origin and propagates at 30’C but stop replicating at 42’C. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3711
    Illegal NheI site found at 2425
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 3717
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3711
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 3711
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 3711
    Plasmid lacks a suffix.
    Illegal XbaI site found at 3726
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NgoMIV site found at 2319
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.


[edit]
Categories
//plasmidbackbone/assembly
//plasmidbackbone/copynumber/low
//plasmidbackbone/operation
//plasmidbackbone/version/5
Parameters
chassis
copies~5
mcsBioBrick
originpSC101ts
resistanceK