Part:BBa_K1470006
pMIG
pMIG is a novel and easy way to handle mammalian expression vectors and to create fast and efficiently stable cell lines. It contains Long Terminal Repeats (LTR) for the production of viral vectors. There is no packaging signal or gag, pol, env in pMIG. Thus it can be used in any biosafety level 1 laboratory.
Origin, Standardization and Application
This plasmid was originally created by William Hahn and is available as plasmid 9044 at Addgene, we got his permission to submit pMIG as biobrick. It is a high copy plasmid and contains an Ampicillin resistance.
In order to provide this plasmid as a mammalian expression vector, we removed several EcoRI- and PstI-sites and an IRES. Afterwards we inserted a multiple cloning site, consisting of XhoI, EcoRI, NotI, SalI, PstI, HindHIII and ClaI. However, it was not possible to delete Xba I-sites in 3'- and 5' LTRs due to their nearly identical sequences. Nonetheless, any biobrick cut with EcoRI/PstI can be ligated in pMIG, hence providing future iGEM Teams a well functioning and reliable expression vector for mammalian cells.
Because of pMIG's retroviral origins, it can be used only in packaging cell lines such as Phoenix cells to produce viral vectors. Otherwise there will not be any functional vectors due to the lack of gag, pol and env.No additional promotor is required for expression on the account of 3'LTR acting as one.
The viral vector uses the murine CAT-1 receptor (mCAT-1) as an entry site into the cell. Therefore, it can specifically transduce mouse cells (e.g. NIH3T3), but not cells lines from other species (e.g. the human embryonic kidney (HEK-293T) cells).
Sequence and Features
Plasmid lacks a suffix.
Illegal EcoRI site found at 4917
Illegal XbaI site found at 289
Illegal XbaI site found at 3711
Illegal PstI site found at 10
Plasmid lacks a suffix.
Illegal EcoRI site found at 4917
Illegal NheI site found at 99
Illegal NheI site found at 3521
Illegal PstI site found at 10
Illegal NotI site found at 4925
Plasmid lacks a suffix.
Illegal EcoRI site found at 4917
Illegal BglII site found at 4899
Illegal XhoI site found at 4905
Plasmid lacks a suffix.
Illegal EcoRI site found at 4917
Illegal XbaI site found at 289
Illegal XbaI site found at 3711
Illegal PstI site found at 10
Plasmid lacks a suffix.
Illegal EcoRI site found at 4917
Illegal XbaI site found at 289
Illegal XbaI site found at 3711
Illegal PstI site found at 10
Illegal NgoMIV site found at 3466
Illegal NgoMIV site found at 4123
Plasmid lacks a suffix.
Illegal BsaI site found at 520
Illegal BsaI site found at 541
Illegal BsaI site found at 3964
Illegal BsaI.rc site found at 1901
Illegal BsaI.rc site found at 4030
Illegal BsaI.rc site found at 4645
Illegal SapI site found at 818
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