Part:BBa_K864014

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.