pMSEx-2 (ATCC® 37836)

Applications: expression vectorintegrating vectorshuttle vector  /  Depositors: M Schuermann

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Designations pMSEx-2
Depositors M Schuermann
Biosafety Level 1
Vector Information
Size (kb): 5.5999999046325680
Vector: pMSEx-2 (plasmid)
Promoters: Promoter SV40 early
Construction: pMSE (ATCC 37834)
Marker(s):G418R,ampR,kanR
Construct size (kb): 5.599999904632568
Features: marker(s): ampR, kanR, G418R
promoter: SV40 early
replicon: pMB1
terminator: MSV LTR
enhancer: SV40 early
Applications
expression vector
integrating vector
shuttle vector
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI/HindIII--2.6, 1.7, 1.0, 0.3; HindIII--2.9, 2.7; EcoRI--4.2, 1.4; BglII--5.6.
A EcoRI/HindIII digest gives fragments of 2.63, 1.68, 0.97 and 0.32 kb.
The SV40 promoter has a 3 bp deletion at the BglI site within the origin of replication to prevent autonomous replication in the presence of SV40 large T-antigen.
The sequence of the oligonucleotide is AGCTTGGTAC CATGGACCTC GAGGCCTAGC TAGCTAG GAGCT.
Kanamycin resistance should be checked at regular intervals. This is best done during amplification steps using LB medium supplemented with 50 ug/ml ampicillin and 12.5 ug/ml kanamycin.
One of 3 shuttle expression vectors (pMSEx-1-3, ATCC 37835-37837) containing a synthetic oligonucleotide providing a translation initiation signal, followed by a StuI site in 3 different positions, followed by a stop signal in all reading frames.
The MSV LTR provides a polyadenylation signal as well as an enhancer for the P1/thymidine kinase promoter regulating the Tn5 kanR/G418R transcription unit.
The order of the major features in this plasmid is: SV40 promoter - oligonucleotide/StuI/oligonucleotide - MLV-LTR - P1 promoter - TK promoter - kanR/G418R - TK polyadenylation signal - ampR.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Schuermann M. An expression vector system for stable expression of oncogenes. Nucleic Acids Res. 18: 4945-4946, 1990. PubMed: 2395667

Marcus Schuermann, personal communication

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