Difference between revisions of "PGP382"
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[[File:PGP382_cloning_region.jpeg|250px|thumb|right|'''pGP382 cloning region: click to enlarge''']] | [[File:PGP382_cloning_region.jpeg|250px|thumb|right|'''pGP382 cloning region: click to enlarge''']] | ||
| − | + | '''Application:''' | |
| − | constructed in [[Stülke]] lab | + | The vector was constructed in the [[Stülke]] lab and it is suitable for constitutive overexpression of C-terminally Strep-tagged proteins in ''B. subtilis''. The plasmid confers resistance to ampicillin and erythromycin in ''E. coli'' and ''B. subtilis'', respectively. pGP382 can be used for the [[SPINE]] method. [[pGP380]] is similar to the vector pGP382 which is derived from vectors [[pBQ200]] and [[pHT315]]. A Shine-Dalgarno sequence has to be fused to the open reading frame during PCR. |
| − | + | Sequencing primers: | |
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*'''M13_puc_for:''' 5‘-GTAAAACGACGGCCAGTG-3‘ | *'''M13_puc_for:''' 5‘-GTAAAACGACGGCCAGTG-3‘ | ||
*'''M13_puc_rev:''' 5‘-GGAAACAGCTATGACCATG-3‘ | *'''M13_puc_rev:''' 5‘-GGAAACAGCTATGACCATG-3‘ | ||
| + | *'''FX125 (rev; primes -114bp of MCS):''' 5‘-GGCTCGTATGTTGTGTGG-3‘ | ||
| + | *'''JN54 (fwd; primes +63bp of MCS):''' 5‘-GTGAAAAATGAGCCGAAAGCAG-3‘ | ||
| − | + | <pubmed> 17994626 </pubmed> | |
| − | + | == Return to [[Plasmids]] == | |
Latest revision as of 08:14, 29 July 2014
Application:
The vector was constructed in the Stülke lab and it is suitable for constitutive overexpression of C-terminally Strep-tagged proteins in B. subtilis. The plasmid confers resistance to ampicillin and erythromycin in E. coli and B. subtilis, respectively. pGP382 can be used for the SPINE method. pGP380 is similar to the vector pGP382 which is derived from vectors pBQ200 and pHT315. A Shine-Dalgarno sequence has to be fused to the open reading frame during PCR.
Sequencing primers:
- M13_puc_for: 5‘-GTAAAACGACGGCCAGTG-3‘
- M13_puc_rev: 5‘-GGAAACAGCTATGACCATG-3‘
- FX125 (rev; primes -114bp of MCS): 5‘-GGCTCGTATGTTGTGTGG-3‘
- JN54 (fwd; primes +63bp of MCS): 5‘-GTGAAAAATGAGCCGAAAGCAG-3‘
