Plasmid_Backbone

Part:BBa_K864005

Designed by: Erik Lundin   Group: iGEM12_Uppsala_University   (2012-09-23)

Low copy BioBrick standard vector

pSB4C15(FRT) is a BioBrick standard vector with low copy pSC101 replication origin BBa_K864050 (~5 copies per cell) and ampicillin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. 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.

The resistance cassette is flanked by FRT sites (BBa_J61020) which allows for subsequent removal of the resistance cassette from the chromosome by the FLP recombinase.

Usage and Biology

This part is available as pSB4A15-pUCori-red. This facilitates high plasmid yields and fast red color expression on transformation plates.

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 3513
    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 3519
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3513
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 3513
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 3513
    Plasmid lacks a suffix.
    Illegal XbaI site found at 3528
    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
originpSC101
resistanceC