pLVX-EF1α-IRES-puro

$216.00

  • Model: PVT2308
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pLVX-EF1α-IRES-puro

PVT2308    2ug
 

pLVX-EF1α-IRES-puro Information

Promoter: EF-1 alpha

Replicon: pUC ori

Terminator: SV40 poly (A) signal

Plasmid classification: virus series, lentivirus cloning vector

Plasmid size: 8825bp

Prokaryotic resistance: Amp

Selection marker: Puro

Clone strain: Stbl3

Culture conditions: 37 C, aerobic LB

Expression host: lactation cells

Induction mode: no need to induce, transient expression.

Primers for 5'sequencing: primers designed based on sequences

Primers for 3'sequencing: primers designed based on sequences

 

pLVX-EF1α-IRES-puro Description

pLVX-EF1α-IRES-Puro contains all of the viral processing elements necessary for the production of replicationincompetent lentivirus, as well as elements to improve viral titer, transgene expression, and overall vector function. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and enhances nuclear export of viral RNA, leading to increased viral titers from packaging cells. In addition, the vector includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus. Finally, pLVX-EF1α-IRES-Puro also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, this element creates a central DNA flap that increases nuclear import ofthe viral genome, resulting in improved vector integration and more efficient transduction. The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria.

 

pLVX-EF1α-IRES-puro Multiple cloning site

pLVX-EF1α-IRES-puro-vector

 

 

pLVX-EF1α-IRES-puro Sequence

LOCUS       Exported                8825 bp ds-DNA     circular SYN 27-AUG-2016

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    Untitled 22

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 8825)

  AUTHORS   .

  TITLE     Direct Submission

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

FEATURES             Location/Qualifiers

     source          1..8825

                     /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)"

     promoter        2338..3519

                     /note="EF-1-alpha promoter"

                     /note="strong constitutive promoter for human elongation 

                     factor EF-1-alpha"

     intron          2568..3510

                     /note="EF-1-alpha intron A"

                     /note="intron upstream of the start codon of human 

                     EF-1-alpha"

     misc_feature    3575..4148

                     /note="IRES"

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

                     encephalomyocarditis virus (EMCV)"

     CDS             4181..4780

                     /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    4794..5382

                     /note="WPRE"

                     /note="woodchuck hepatitis virus posttranscriptional 

                     regulatory element"

     CDS             complement(5265..5276)

                     /codon_start=1

                     /product="Factor Xa recognition and cleavage site"

                     /note="Factor Xa site"

                     /translation="IEGR"

     LTR             5589..6222

                     /note="3' LTR"

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

     primer_bind     complement(6350..6366)

                     /note="M13 rev"

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

                     variants"

     protein_bind    6374..6390

                     /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(6398..6428)

                     /note="lac promoter"

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

     protein_bind    6443..6464

                     /bound_moiety="E. coli catabolite activator protein"

                     /note="CAP binding site"

                     /note="CAP binding activates transcription in the presence 

                     of cAMP."

     rep_origin      complement(6752..7340)

                     /direction=LEFT

                     /note="ori"

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

                     replication"

     CDS             complement(7511..8371)

                     /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(8372..8476)

                     /gene="bla"

                     /note="AmpR promoter"

     polyA_signal    8524..8658

                     /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 cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg

     2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc

     2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct

     2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag

     2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg

     2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag

     2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt

     2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt

     2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg

     2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg

     2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct

     2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg

     2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc

     2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga

     3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct

     3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca

     3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg

     3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg

     3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat

     3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg

     3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa

     3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca

     3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gatctatttc cggtgaattc

     3541 ctcgagacta gttctagagc ggccgcggat cccgcccctc tccctccccc ccccctaacg

     3601 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca

     3661 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga

     3721 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga

     3781 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca

     3841 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag

     3901 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa

     3961 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac

     4021 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga

     4081 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac

     4141 gatgataagc ttgccacaac ccacaaggag acgaccttcc atgaccgagt acaagcccac

     4201 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt

     4261 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac

     4321 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc

     4381 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt

     4441 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca

     4501 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac

     4561 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg

     4621 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa

     4681 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg

     4741 accgcgcacc tggtgcatga cccgcaagcc cggtgcctag acgcgtctgg aacaatcaac

     4801 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta

     4861 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt

     4921 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg

     4981 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg

     5041 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca

     5101 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca

     5161 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg

     5221 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag

     5281 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc

     5341 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag

     5401 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga

     5461 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt

     5521 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa

     5581 gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat

     5641 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag

     5701 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga

     5761 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga

     5821 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt

     5881 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg

     5941 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc

     6001 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc

     6061 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa

     6121 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga

     6181 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc

     6241 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac

     6301 attgctagcg tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt

     6361 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa

     6421 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac

     6481 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg

     6541 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc

     6601 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat

     6661 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca

     6721 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc

     6781 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc

     6841 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg

     6901 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta

     6961 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg

     7021 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac

     7081 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag

     7141 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat

     7201 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat

     7261 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc

     7321 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt

     7381 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct

     7441 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt

     7501 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc

     7561 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac

     7621 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat

     7681 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg

     7741 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata

     7801 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta

     7861 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt

     7921 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag

     7981 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa

     8041 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc

     8101 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt

     8161 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc

     8221 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta

     8281 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa

     8341 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca

     8401 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac

     8461 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag

     8521 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc

     8581 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta

     8641 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc

     8701 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat

     8761 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta

     8821 gtagt

//

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