Basic Vector Information
- Vector Name:
- pMSCV PIG (Puro IRES GFP empty vector)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7627 bp
- Type:
- Mammalian Expression, Retroviral
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- High Copy
- Promoter:
- MSCV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- pBABE 5
- 3' Primer:
- EGFP-N
pMSCV PIG (Puro IRES GFP empty vector) vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pMSCV PIG (Puro IRES GFP empty vector) vector Sequence
LOCUS 40924_32380 7627 bp DNA circular SYN 16-AUG-2021 DEFINITION Retroviral backbone for cloning and gene expression. Select with puromycin or GFP.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7627) AUTHORS Mayr C, Bartel DP TITLE Widespread shortening of 3'UTRs by alternative cleavage and polyadenylation activates oncogenes in cancer cells. JOURNAL Cell. 2009 Aug 21. 138(4):673-84. PUBMED 19703394 REFERENCE 2 (bases 1 to 7627) TITLE Direct Submission REFERENCE 3 (bases 1 to 7627) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2009 Aug 21. 138(4):673-84." COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..7627 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind complement(676..692) /label=lac operator /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(700..730) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(745..766) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(883..900) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1054..1642) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(1816..2673) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(2674..2778) /label=AmpR promoter primer_bind 2846..2864 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(2902..2924) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3024..3043 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3237..3259 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3387..3406 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 3606..4122 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" misc_feature 4186..4527 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 4594..5010 /codon_start=1 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" /translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA" promoter 5045..5544 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" CDS 5565..6161 /codon_start=1 /label=PuroR /note="puromycin N-acetyltransferase" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" misc_feature 6229..6805 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 6806..7522 /codon_start=1 /label=EGFP /note="enhanced GFP" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" LTR 7627 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus"
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