pLVX- IRES- Zsgreen1 Plasmid

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  • Model: PVT2305
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pLVX- IRES- Zsgreen1 Plasmid

PVT2305      2ug

 

pLVX-IRES-ZsGreen1 Information

Promoter: CMV
Replicon: pUC ori
Terminator: WPRE, 3 'LTR
Plasmid classification: virus series, lentivirus cloning vector
Plasmid size: 8204bp
Prokaryotic resistance: Amp
Cloned strain: Stbl3
Culture conditions: 37 centigrade, aerobic LB
Expression host: mammalian cells
Induction mode: no induction, instantaneous expression
5'sequencing primers: CMV-F:CGCAAATGGGCGGTAGGCGTG
3'sequencing primers: primers designed according to sequence
Use:Lentiviral

 

pLVX-IRES-ZsGreen1 Description

Plvx-ires-zsgreen1 is HIV-1, lentiviral expression vector, which enables you to express interest in almost any mammalian cell type and zsgreen1 protein at the same time, including primary cells. Zsgreen1 is a variant of the human codon optimization of coral reef Zoanthus, a green fluorescent protein, ZsGreen. The vector expression of the two protein bis CIS mRNA transcription, making zsgreen1 as an indicator of transduction efficiency, using flow cytometry to select markers.
The transcription of the transcriptional expression of the paramegon activates the immediate early promoter drive (CMV) of the human cytomegalovirus (HCMV) in the upstream MCS. The ribosomal entry site (IRES) of EMCV is located between MCS and zsgreen1, which is conducive to independent translation of zsgreen1 cap in IRES / zsgreen1 junction.
Plvx-ires-zsgreen1 contains all the elements that are necessary for replication of lentivirus, and elements to improve virus titer, expression level of foreign gene, and the whole vector function. The posttranscriptional regulator element (WPRE) of the groundhog hepatitis virus (HBV) promotes RNA treatment events and increases the RNA nuclear exits of the virus, resulting in increased virus titer packaging cells. In addition, the carrier includes the Rev response element (RRE), which further increases the titer of the virus to increase the spliced virus RNA to transport the nucleus. Finally, plvx-ires-zsgreen1 also contains a central purine channel / central termination sequence (CPPT / CTS). In the process of target cell infection, the element creates a central DNA flap, which increases the nuclear import of viral genome, thus improving vector integration and more effective transduction. The carrier also contains a copy of the city Bureau origin and the Escherichia coli resistance gene of ampicillin (Ampr) in bacterial reproduction and selection.

pLVX-IRES-Zsgreen1-plasmid

 

pLVX-IRES-ZsGreen1 Sequence

LOCUS       Exported                8204 bp ds-DNA     circular SYN 27-AUG-2016

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    Untitled 19

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 8204)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Saturday, August 27, 2016 from SnapGene Viewer 3.1.4

FEATURES             Location/Qualifiers

     source          1..8204

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     LTR             1..634

                     /note="3' LTR"

                     /note="3' long terminal repeat (LTR) from HIV-1"

     misc_feature    681..806

                     /note="HIV-1 Psi"

                     /note="packaging signal of human immunodeficiency virus 

                     type 1"

     misc_feature    1303..1536

                     /note="RRE"

                     /note="The Rev response element (RRE) of HIV-1 allows for 

                     Rev-dependent mRNA export from the nucleus to the 

                     cytoplasm."

     misc_feature    2028..2143

                     /note="cPPT/CTS"

                     /note="central polypurine tract and central termination 

                     sequence of HIV-1 (lacking the first T)"

     enhancer        2201..2504

                     /note="CMV enhancer"

                     /note="human cytomegalovirus immediate early enhancer"

     promoter        2505..2708

                     /note="CMV promoter"

                     /note="human cytomegalovirus (CMV) immediate early 

                     promoter"

     misc_feature    2843..3415

                     /note="IRES"

                     /note="internal ribosome entry site (IRES) of the 

                     encephalomyocarditis virus (EMCV)"

     misc_feature    2866..3417

                     /note="IRES"

                     /note="internal ribosome entry site (IRES) of the 

                     encephalomyocarditis virus (EMCV)"

     CDS             3417..4112

                     /codon_start=1

                     /product="Zoanthus green fluorescent protein"

                     /note="ZsGreen1"

                     /note="mammalian codon-optimized"

                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN

                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV

                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG

                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH

                     AIASGSALP"

     misc_feature    4126..4714

                     /note="WPRE"

                     /note="woodchuck hepatitis virus posttranscriptional 

                     regulatory element"

     CDS             complement(4597..4608)

                     /codon_start=1

                     /product="Factor Xa recognition and cleavage site"

                     /note="Factor Xa site"

                     /translation="IEGR"

     LTR             4921..5554

                     /note="3' LTR"

                     /note="3' long terminal repeat (LTR) from HIV-1"

     primer_bind     complement(5682..5698)

                     /note="M13 rev"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     protein_bind    5706..5722

                     /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(5730..5760)

                     /note="lac promoter"

                     /note="promoter for the E. coli lac operon"

     protein_bind    5775..5796

                     /bound_moiety="E. coli catabolite activator protein"

                     /note="CAP binding site"

                     /note="CAP binding activates transcription in the presence 

                     of cAMP."

     rep_origin      complement(6084..6672)

                     /direction=LEFT

                     /note="ori"

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

     CDS             complement(6843..7703)

                     /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(7704..7808)

                     /gene="bla"

                     /note="AmpR promoter"

     polyA_signal    7856..7990

                     /note="SV40 poly(A) signal"

                     /note="SV40 polyadenylation signal"

ORIGIN

        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca

       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac

      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca

      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg

      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag

      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg

      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat

      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga

      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct

      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc

      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag

      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg

      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga

      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg

      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg

      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct

      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat

     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca

     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc

     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag

     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc

     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg

     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc

     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc

     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct

     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa

     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca

     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt

     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt

     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta

     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt

     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct

     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga

     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag

     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac

     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg

     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa

     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg

     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt

     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg

     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc

     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc

     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca

     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta

     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa

     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc

     2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcct cgagactagt

     2821 tctagagcgg ccgcggatcc cgcccctctc cctccccccc ccctaacgtt actggccgaa

     2881 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt

     2941 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg

     3001 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc

     3061 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc

     3121 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa

     3181 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc

     3241 tcacctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg

     3301 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg

     3361 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg

     3421 cccagtccaa gcacggcctg accaaggaga tgaccatgaa gtaccgcatg gagggctgcg

     3481 tggacggcca caagttcgtg atcaccggcg agggcatcgg ctaccccttc aagggcaagc

     3541 aggccatcaa cctgtgcgtg gtggagggcg gccccttgcc cttcgccgag gacatcttgt

     3601 ccgccgcctt catgtacggc aaccgcgtgt tcaccgagta cccccaggac atcgtcgact

     3661 acttcaagaa ctcctgcccc gccggctaca cctgggaccg ctccttcctg ttcgaggacg

     3721 gcgccgtgtg catctgcaac gccgacatca ccgtgagcgt ggaggagaac tgcatgtacc

     3781 acgagtccaa gttctacggc gtgaacttcc ccgccgacgg ccccgtgatg aagaagatga

     3841 ccgacaactg ggagccctcc tgcgagaaga tcatccccgt gcccaagcag ggcatcttga

     3901 agggcgacgt gagcatgtac ctgctgctga aggacggtgg ccgcttgcgc tgccagttcg

     3961 acaccgtgta caaggccaag tccgtgcccc gcaagatgcc cgactggcac ttcatccagc

     4021 acaagctgac ccgcgaggac cgcagcgacg ccaagaacca gaagtggcac ctgaccgagc

     4081 acgccatcgc ctccggctcc gccttgccct gaacgcgtct ggaacaatca acctctggat

     4141 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt

     4201 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc

     4261 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg

     4321 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc

     4381 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa

     4441 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat

     4501 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc

     4561 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt

     4621 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag

     4681 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc

     4741 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct

     4801 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa

     4861 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac

     4921 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca

     4981 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac

     5041 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca

     5101 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg

     5161 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag

     5221 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg

     5281 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat

     5341 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga

     5401 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct

     5461 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc

     5521 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt

     5581 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag

     5641 cgtttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg

     5701 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa

     5761 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct

     5821 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga

     5881 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc

     5941 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa

     6001 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt

     6061 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa

     6121 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt

     6181 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg

     6241 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc

     6301 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc

     6361 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta

     6421 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct

     6481 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc

     6541 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa

     6601 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa

     6661 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa

     6721 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt

     6781 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac

     6841 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc

     6901 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc

     6961 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata

     7021 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc

     7081 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc

     7141 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca

     7201 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa

     7261 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca

     7321 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt

     7381 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt

     7441 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg

     7501 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga

     7561 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc

     7621 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg

     7681 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag

     7741 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg

     7801 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt

     7861 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa

     7921 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca

     7981 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata

     8041 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga

     8101 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt

     8161 taaagaaatt gtatttgtta aatatgtact acaaacttag tagt

//


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pLVX-IRES-Zsgreen1,Plasmid pLVX-IRES-Zsgreen1,pLVX-IRES-Zsgreen1 vector

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