Basic Vector Information
- Vector Name:
- pMSCV-Zeo
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6017 bp
- Type:
- Mammalian Expression, Retroviral
- Replication origin:
- ori
- Selection Marker:
- Zeocin
- Promoter:
- MSCV
- Cloning Method:
- Restriction Enzyme
pMSCV-Zeo 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-Zeo vector Sequence
LOCUS 40924_32355 6017 bp DNA circular SYN 03-DEC-2021
DEFINITION retrovirus-mediated gene transfer into mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6017)
AUTHORS Kendall J, Liu Q, Bakleh A, Krasnitz A, Nguyen KC, Lakshmi B, Gerald
WL, Powers S, Mu D
TITLE Oncogenic cooperation and coamplification of developmental
transcription factor genes in lung cancer.
JOURNAL Proc Natl Acad Sci U S A. 2007 Oct 16;104(42):16663-8. Epub 2007 Oct
9.
PUBMED 17925434
REFERENCE 2 (bases 1 to 6017)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6017)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
Acad Sci U S A."; date: "2007-10-16"; volume: "104(42)"; pages:
"16663-8. Epub 2007 Oct 9"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6017
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..515
/label=MSCV
/note="Murine embryonic stem cell virus promoter including
the enhancer and promoter region of PCMV virus and 5'
untranslated region of dl-587rev retrovirus"
misc_feature 579..920
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 987..1403
/codon_start=1
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
/translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
promoter 1438..1937
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 1958..2329
/codon_start=1
/label=BleoR
/note="antibiotic-binding protein"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
LTR 2379..2893
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
primer_bind complement(3062..3078)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3086..3102)
/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(3110..3140)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3155..3176)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3293..3310)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3464..4052)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4226..5083)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5084..5188)
/label=AmpR promoter
primer_bind 5256..5274
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(5312..5334)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 5434..5453
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5647..5669
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 5797..5816
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
LTR 6016..6017
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
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