Difference between revisions of "Part:BBa K864014"
 
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<partinfo>BBa_K864014 short</partinfo>
 
<partinfo>BBa_K864014 short</partinfo>
  
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pSB4K15Iq(FRT) is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and kanamycin 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.
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pSB4K15Iq(FRT) is a BioBrick standard vector with low copy pSC101 replication origin <partinfo>BBa_K864050</partinfo> (~5 copies per cell) and kanamycin 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.
  
 
This vector carries a LacIq cassette on the backbone for tight repression of any lac promoter cloned into the cloning site.
 
This vector carries a LacIq cassette on the backbone for tight repression of any lac promoter cloned into the cloning site.
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The resistance cassette is flanked by FRT sites ([[Part:BBa_J61020|BBa_J61020]]) which allows for subsequent removal of the resistance cassette from the chromosome by the FLP recombinase.  
 
The resistance cassette is flanked by FRT sites ([[Part:BBa_J61020|BBa_J61020]]) which allows for subsequent removal of the resistance cassette from the chromosome by the FLP recombinase.  
  
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===Usage and Biology===
 
===Usage and Biology===
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This part is available as pSB4A15-[[Part:BBa_K864121|pUCori-redq]]. This facilitates high plasmid yields and fast red color expression on transformation plates.
  
 
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Latest revision as of 19:23, 27 September 2012

Low copy BioBrick standard vector with LacI

pSB4K15Iq(FRT) is a BioBrick standard vector with low copy pSC101 replication origin BBa_K864050 (~5 copies per cell) and kanamycin 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.

This vector carries a LacIq cassette on the backbone for tight repression of any lac promoter cloned into the cloning site.

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-redq. 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 4891
    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 4897
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 4891
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 4891
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 4891
    Plasmid lacks a suffix.
    Illegal XbaI site found at 4906
    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.