pLVX-IRES-Neo

Catalog No. PVT11065
Packing 2ug

 

pLVX-IRES-Neo information

Promoter: CMV

Replicon: pUC ori

Terminator: SV40 poly (A) signal

Plasmid classification: virus series, lentivirus cloning vector

Plasmid size: 8269bp

Prokaryotic resistance: Amp

Screening markers: Neo

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-Neo Description

pLVX-IRES-Neo is an HIV-1-based, lentiviral expression vector that allows the simultaneous expression of your protein of interest and G418 resistance (Neor) in virtually any mammalian cell type, including primary cells. The vector expresses your protein of interest and Neor from a bicistronic mRNA transcript, allowing G418 resistance to be used as an indicator of transduction efficiency and a marker for selection. pLVX-IRES-Neo 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-Neo 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-Neo Sequence

LOCUS       Exported                8269 bp ds-DNA     circular SYN 27-AUG-2016

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    Untitled 20

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 8269)

  AUTHORS   .

  TITLE     Direct Submission

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

FEATURES             Location/Qualifiers

     source          1..8269

                     /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             3430..4224

                     /codon_start=1

                     /gene="aph(3')-II (or nptII)"

                     /product="aminoglycoside phosphotransferase from Tn5"

                     /note="NeoR/KanR"

                     /note="confers resistance to neomycin, kanamycin, and G418 

                     (Geneticin(R))"

                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP

                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS

                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ

                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA

                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"

     misc_feature    4238..4826

                     /note="WPRE"

                     /note="woodchuck hepatitis virus posttranscriptional 

                     regulatory element"

     CDS             complement(4709..4720)

                     /codon_start=1

                     /product="Factor Xa recognition and cleavage site"

                     /note="Factor Xa site"

                     /translation="IEGR"

     LTR             5033..5666

                     /note="3' LTR"

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

     primer_bind     complement(5794..5810)

                     /note="M13 rev"

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

                     variants"

     protein_bind    5818..5834

                     /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(5842..5872)

                     /note="lac promoter"

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

     protein_bind    5887..5908

                     /bound_moiety="E. coli catabolite activator protein"

                     /note="CAP binding site"

                     /note="CAP binding activates transcription in the presence 

                     of cAMP."

     rep_origin      complement(6196..6784)

                     /direction=LEFT

                     /note="ori"

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

                     replication"

     CDS             complement(6955..7815)

                     /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(7816..7920)

                     /gene="bla"

                     /note="AmpR promoter"

     polyA_signal    7968..8102

                     /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 gccacaacca tggctgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag

     3481 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc

     3541 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg

     3601 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc

     3661 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg

     3721 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct

     3781 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg

     3841 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat

     3901 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc

     3961 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg

     4021 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc

     4081 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct

     4141 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat

     4201 cgccttcttg acgagttctt ctgaacgcgt ctggaacaat caacctctgg attacaaaat

     4261 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc

     4321 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt

     4381 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg

     4441 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg

     4501 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc

     4561 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt

     4621 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct

     4681 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg

     4741 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg

     4801 gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa

     4861 catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa

     4921 gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg

     4981 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg

     5041 ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc

     5101 tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt

     5161 ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc caataaagga

     5221 gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa

     5281 gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat

     5341 ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac

     5401 tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata

     5461 taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga

     5521 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct

     5581 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt

     5641 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt

     5701 ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgct agcgtttacc

     5761 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg

     5821 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg

     5881 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc

     5941 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt

     6001 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct

     6061 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga

     6121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc

     6181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg

     6241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg

     6301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt

     6361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt

     6421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg

     6481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact

     6541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt

     6601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct

     6661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac

     6721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc

     6781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg

     6841 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta

     6901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca

     6961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc

     7021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc

     7081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc

     7141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat

     7201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt

     7261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc

     7321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag

     7381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt

     7441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac

     7501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg

     7561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat

     7621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc

     7681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc

     7741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa

     7801 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg

     7861 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg

     7921 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac ttgtttattg

     7981 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt

     8041 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga

     8101 tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta

     8161 ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt

     8221 cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagt

//

 

Caution:
1.  This product is FOR RESEARCH USE ONLY!
2.  The item is lyophilized form, Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid.