Part:BBa_K4382015

Partial Sequence of pCri-18a plasmid with additional SalI site

This is the specific sequence of pCri-18a plasmid backbone which contains the additional SalI site.The sequence of the MCS of the plasmid is available online.

Usage and Biology

The pCri system is a vector collection for cytoplasmic and periplasmic/extracellular expression of heterologous proteins that allows the simultaneous assessment of prokaryotic and eukaryotic host cells (Escherichia coli, Bacillus subtilis, and Pichia pastoris). By using a single polymerase chain reaction product, genes of interest can be directionally cloned in all vectors within four different rare restriction sites at the 5′ end and multiple cloning sites at the 3′end.

The pCri System allows the fusion of a His6–8-tag at the N-terminus of the target protein, which can be in tandem with larger tags such MBP , GST , small ubiquitin-like modifier (SUMO), and the β-trefoil lectin module of protein LSL150 from the mushroom Laetiporus sulphureus (LSL). The C-terminus of the target protein can likewise be furnished with a His6-tag or a Strep-tag if the stop codon of the amplified gene is omitted. These tags add a functionality to the target protein, which is commonly used as a first purification step through affinity chromatography.

Total vector size (bp): ~ 8700

Vector type: Shuttle vector for E.coli and Bacillus Subtilis

Bacterial Resistance(s): Ampicillin, (100 μg/mL) for E.coli, Chloramphenicol (5 μg/mL) for Bacillus Subtilis

Growth Temperature: 37°C

Growth Strain(s): DH5alpha

Copy number: High Copy

References

1)Goulas, T., Cuppari, A., Garcia-Castellanos, R., Snipas, S., Glockshuber, R., Arolas, J. L., & Gomis-Rüth, F. X. (2014, November 11). The pCri System: A Vector Collection for Recombinant Protein Expression and Purification. PLoS ONE, 9(11), e112643. https://doi.org/10.1371/journal.pone.0112643

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.