pLVX- IRES- Puro

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  • Model: PVT2304
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pLVX-IRES-Puro

PVT2304      2ug
 

pLVX-IRES-Puro plasmid information

Promoter: CMV
Replicon: pUC ori
Terminator: SV40 poly (A) signal
Plasmid classification: virus series, lentivirus cloning vector
Plasmid size: 8140bp
Prokaryotic resistance: Amp
Screening markers: Puro
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-Puro-plasmid

pLVX-IRES-Puro plasmid Description
pLVX-IRES-Puro is an HIV-1-based, lentiviral expression vector that allows the simultaneous expression of your protein of interest and puromycin resistance (Puror) in virtually any mammalian cell type, including primary cells. The vector expresses your protein of interest and Puror from a bicistronic mRNA transcript, allowing puromycin resistance to be used as an indicator of transduction efficiency and a marker for selection. Expression of the bicistronic transcript is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE) located just upstream of the MCS. An encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES), positioned between the MCS and Puror , facilitates cap-independent translation of Puror from an internal start site at the IRES/Puror junction . pLVX-IRES-Puro contains all of the viral processing elements necessary for the production of replication-incompetent 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-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 of the 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-IRES-Puro plasmid Sequence
 

LOCUS       Exported                8140 bp ds-DNA     circular SYN 27-AUG-2016

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    Untitled 18

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 8140)

  AUTHORS   .

  TITLE     Direct Submission

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

FEATURES             Location/Qualifiers

     source          1..8140

                     /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..3416

                     /note="IRES"

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

                     encephalomyocarditis virus (EMCV)"

     CDS             3449..4048

                     /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    4062..4650

                     /note="WPRE"

                     /note="woodchuck hepatitis virus posttranscriptional 

                     regulatory element"

     CDS             complement(4533..4544)

                     /codon_start=1

                     /product="Factor Xa recognition and cleavage site"

                     /note="Factor Xa site"

                     /translation="IEGR"

     LTR             4857..5490

                     /note="3' LTR"

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

     primer_bind     complement(5618..5634)

                     /note="M13 rev"

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

                     variants"

     protein_bind    5642..5658

                     /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(5666..5696)

                     /note="lac promoter"

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

     protein_bind    5711..5732

                     /bound_moiety="E. coli catabolite activator protein"

                     /note="CAP binding site"

                     /note="CAP binding activates transcription in the presence 

                     of cAMP."

     rep_origin      complement(6020..6608)

                     /direction=LEFT

                     /note="ori"

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

                     replication"

     CDS             complement(6779..7639)

                     /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(7640..7744)

                     /gene="bla"

                     /note="AmpR promoter"

     polyA_signal    7792..7926

                     /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 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg

     3301 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac

     3361 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt

     3421 gccacaaccc acaaggagac gaccttccat gaccgagtac aagcccacgg tgcgcctcgc

     3481 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc

     3541 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga

     3601 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc

     3661 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat

     3721 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg

     3781 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc

     3841 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc

     3901 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta

     3961 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg

     4021 gtgcatgacc cgcaagcccg gtgcctagac gcgtctggaa caatcaacct ctggattaca

     4081 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat

     4141 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct

     4201 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac

     4261 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca

     4321 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca

     4381 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg

     4441 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga

     4501 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt

     4561 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga

     4621 gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc gagacctaga

     4681 aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct

     4741 agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc

     4801 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaaga ggggactgga

     4861 agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca

     4921 aggctacttc cctgattagc agaactacac accagggcca ggggtcagat atccactgac

     4981 ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa

     5041 aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag

     5101 agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct

     5161 gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg

     5221 ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg

     5281 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct

     5341 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag

     5401 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac

     5461 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt

     5521 atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat tgctagcgtt

     5581 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa

     5641 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct

     5701 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc

     5761 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg

     5821 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc

     5881 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag

     5941 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa

     6001 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc

     6061 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc

     6121 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg

     6181 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt

     6241 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc

     6301 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc

     6361 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag

     6421 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg

     6481 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa

     6541 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag

     6601 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact

     6661 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa

     6721 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt

     6781 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag

     6841 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca

     6901 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc

     6961 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt

     7021 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg

     7081 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca

     7141 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg

     7201 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca

     7261 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg

     7321 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct

     7381 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca

     7441 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca

     7501 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg

     7561 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac

     7621 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt

     7681 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc

     7741 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt

     7801 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca

     7861 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc

     7921 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct

     7981 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta

     8041 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agtttttaaa

     8101 gaaattgtat ttgttaaata tgtactacaa acttagtagt

//

Caution:
Product is for research use only!

 

Search name

pLVX-IRES-Puro,Plasmid pLVX-IRES-Puro,pLVX-IRES-Puro vector

 

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