Part:BBa_K4417014

pCT5c backbone

Description

This part pCT5c backbone (BBa_K4417014) is a plasmid backbone based on pCT5c (BBa_K4417000). This part was digested from the urease master plasmid (BBa_K4417017). The backbone is flanked with SapI Type IIS prefix and suffix, enabling Type IIS cloning. The SapI prefix and suffix are setting off the assembly compatibility detector below.

Figure 1: pCT5c backbone with SapI prefix and suffix. SapI binding sites were shaded in grey, and fusion sites were in red.

Usage and Biology

• This part can be used in Type IIS restriction cloning.
• E. coli ori is a pMB1 derivative.
• B. sub ori is unknown.
• Two antibiotic selections, with ampicillin under E. coli promoter and B. subtilis under tetracycline promoter.

Reference

1. Development of a synthetic cumate-inducible gene expression system for Bacillus. Seo SO, Schmidt-Dannert C. Appl Microbiol Biotechnol. 2018 Nov 3. pii: 10.1007/s00253-018-9485-4. doi: 10.1007/s00253-018-9485-4. 10.1007/s00253-018-9485-4 PubMed 30392122

2. Addgene plasmid # 119872; http://n2t.net/addgene:119872; RRID:Addgene_119872

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2009
  Illegal EcoRI site found at 5065
  Illegal XbaI site found at 5520
  Illegal SpeI site found at 5961
  Illegal PstI site found at 1257
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2009
  Illegal EcoRI site found at 5065
  Illegal NheI site found at 5296
  Illegal SpeI site found at 5961
  Illegal PstI site found at 1257
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2009
  Illegal EcoRI site found at 5065
  Illegal BamHI site found at 6867
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2009
  Illegal EcoRI site found at 5065
  Illegal XbaI site found at 5520
  Illegal SpeI site found at 5961
  Illegal PstI site found at 1257
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2009
  Illegal EcoRI site found at 5065
  Illegal XbaI site found at 5520
  Illegal SpeI site found at 5961
  Illegal PstI site found at 1257
  Illegal NgoMIV site found at 20
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal SapI site found at 6873
  Illegal SapI.rc site found at 5