pET His10 MBP Asn10 TEV LIC cloning vector (2CT-10)
(Plasmid #55209)

• Purpose: (Empty Backbone)
• Depositing Lab: Scott Gradia
• Publication: 2-series LIC N-terminal fusion vectors for E. coli expression (unpublished)

Backbone

• Vector backbone: pET
• Vector type: Bacterial Expression
• Tags / Fusion Proteins:
  • His10 (N terminal on backbone)
  • MPB, Asn10 (N terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: Low Copy

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: T7 Forward
• 3′ sequencing primer: T7 reverse

Resource Information

• Articles Citing this Plasmid:
  • 8 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This plasmid is an empty vector. Your gene can be inserted with a LIC cloning protocol. All 2-series vectors work as single-expression vectors, as well as transfer vectors for our polycistronic system.

The LIC cloning site is flanked by 5 pairs of restriction sites, so that your gene can easily be subcloned into our polycistronic destination vectors (2D, 2E, or 2Z).

2CT-10 has a TEV-cleavable N-terminal His10-MBP fusion tag followed by Asn10. MBP can improve the expression and solubility of your target protein. The N10 linker may help you to avoid steric clashes between your protein and the MBP tag.

To clone into this vector, add LIC v1 tags to the 5' end of your PCR primers.

Forward - 5'-TACTTCCAATCCAATGCA-3'
Reverse - 5'-TTATCCACTTCCAATGTTATTA-3'

Linearize the plasmid with SspI and gel purify.

When digesting the DNA with T4 polymerase, use dCTP for insert and dGTP for vector.