pMSCVpuro-IRES-EGFP

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  • Model: PVT11051
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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

     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag

     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc

     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc

     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg

     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag

     1381 ccctcactcc ttctctaggc gccggaatta gatctctcga ggttaacgaa ttctaccggg

     1441 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca

     1501 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc

     1561 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga

     1621 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc

     1681 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg

     1741 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg

     1801 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg

     1861 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct

     1921 catctccggg cctttcgacc tgcagcccaa gcttaccatg accgagtaca agcccacggt

     1981 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc

     2041 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga

     2101 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga

     2161 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt

     2221 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca

     2281 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt

     2341 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt

     2401 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct

     2461 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc

     2521 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg

     2581 cccgaccgaa aggagcgcac gaccccatgc atccaattcc gccccccccc cctaacgtta

     2641 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca

     2701 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca

     2761 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg

     2821 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc

     2881 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata

     2941 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag

     3001 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc

     3061 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt

     3121 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat

     3181 gataatatgg ccacaaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc

     3241 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc tggcgagggc

     3301 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg

     3361 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc

     3421 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc

     3481 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag

     3541 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac

     3601 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg

     3661 gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa catcgaggac

     3721 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg

     3781 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag

     3841 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg

     3901 gacgagctgt acaagtaatg aattaattaa gaattatcaa gcttatcgat accgtcgacc

     3961 tgcagccaag cttatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa

     4021 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat

     4081 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca

     4141 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga

     4201 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt

     4261 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc

     4321 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc

     4381 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata

     4441 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg

     4501 agtgattgac tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga

     4561 acaacattct gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt

     4621 tgaatccaca tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg

     4681 gggagaattg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat

     4741 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc

     4801 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg

     4861 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct

     4921 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt

     4981 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc

     5041 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga

     5101 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata

     5161 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac

     5221 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg

     5281 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc

     5341 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag

     5401 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt

     5461 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt

     5521 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg

     5581 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac

     5641 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca

     5701 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac

     5761 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac

     5821 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt

     5881 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt

     5941 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt

     6001 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc

     6061 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa

     6121 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg

     6181 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt

     6241 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc

     6301 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt

     6361 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg

     6421 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac

     6481 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc

     6541 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt

     6601 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg

     6661 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag

     6721 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa

     6781 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat

     6841 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg

     6901 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg

     6961 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg

     7021 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat

     7081 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc

     7141 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca

     7201 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca

     7261 gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac

     7321 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg

     7381 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca

     7441 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt

     7501 gttaaagaca ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg

     7561 aagcctatag agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag

     7621 gggggaa

//


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