pMSCVpuro-IRES-EGFP
Catalog No. PVT11051
Packing 2ug
pMSCVpuro-IRES-EGFP Information
Replicon: pUC ori
Plasmid classification: virus series, reverse virus cloning cloning vector
Prokaryotic resistance: ampicillin Amp
Screening marker: puromycin Puro
Clone strain: Escherichia coli Stbl3
Culture conditions: 37 C, aerobic, LB
Expression host: lactation cells
Induction mode: no need to induce, transient expression, or stable expression.
Primers for 5'sequencing: primers designed based on sequences
Primers for 3'sequencing: primers designed based on sequences
pMSCVpuro-IRES-EGFP Description
Mouse stem cell virus (MSCV) vector is a retroviral vector derived from mouse embryonic stem cell virus (MesV) and LN. In transfected packaging cell lines, pmscv Puro is expressed instantaneously and stably, or integrates. The transcript contains extended viral packaging signal_+, purinomycin resistance genes and target genes. Vectors achieve stable, high-level gene expression in hematopoietic and embryonic stem cells through a specially designed 5 "long-terminal repeat (LTR). This sequence is a virus from the mouse stem cell gene, which differs from several point mutations in other retroviral vectors MoMuLV LTR and deletions. These changes enhance transcriptional activation and inhibit transcriptional inhibition of embryonic stem cells and embryonic cancer cells. As a result, LTR drives the constitutive expression of target genes at higher levels in stem cells or other mammalian cell lines. A polyclonal site where a gene can be cloned, the LTR. Puromycin resistant gene puror downstream of the mouse phosphoglycerol kinase (PKG) promoter, selects antibiotics in eukaryotic cells. Pmscvpuro also contains the origin of replication and Escherichia coli ampR genes in bacterial breeding and antibiotic selection.
pMSCVpuro-IRES-EGFP Sequence
LOCUS Exported 7627 bp ds-DNA circular SYN 27-APR-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..7627
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 579..920
/note="MESV Psi"
/note="packaging signal of murine embryonic stem cell
virus"
CDS 987..1403
/note="gag (truncated)"
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
promoter 1438..1937
/note="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 1958..2557
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="PuroR"
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 2635..3187
/note="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 3199..3918
/codon_start=1
/product="enhanced GFP"
/note="EGFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
LTR 4020..4534
/note="3' LTR"
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind 4696..4712
/bound_moiety="lac repressor encoded by lacI"
/note="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(4720..4750)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4765..4786
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5074..5662)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5833..6693)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6694..6798)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
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