LOCUS       sequence-354458-        8738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8738)
  AUTHORS   Hayer A, Shao L, Chung M, Joubert LM, Yang HW, Tsai FC, Bisaria A, 
            Betzig E, Meyer T
  TITLE     Engulfed cadherin fingers are polarized junctional structures 
            between collectively migrating endothelial cells.
  JOURNAL   Nat Cell Biol. 2016 Dec;18(12):1311-1323. doi: 10.1038/ncb3438. Epub
            2016 Nov 14.
  PUBMED    27842057
REFERENCE   2  (bases 1 to 8738)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/ncb3438"; 
            journalName: "Nat Cell Biol"; date: "2016-12"; volume: "18"; issue: 
            "12"; pages: "1311-1323"
COMMENT     Sequence Label: pLV-EF1a-IRES-Puro
FEATURES             Location/Qualifiers
     source          1..8738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(629..645)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(629..645)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(642..664)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    653..669
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(join(677..683,684..701,702..707))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    722..743
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(860..877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1031..1619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1111..1130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1790..2581,2582..2650))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2413..2432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2651..2755)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2803..2937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2840..2859)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2894..2913
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             3105..3738
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3785..3910
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4407..4640
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4825..4869
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5132..5247
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        5325..5536
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          5537..6475
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6432..6452
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    6636..7185
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6780..6797)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7007..7026
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7188..7787
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7188..7207)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7684..7704
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7811..8399
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7864..7884)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8282..8293)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             join(8606..8738,1..501)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa
       61 caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt
      121 agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga
      181 gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc
      241 agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc
      301 agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct
      361 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
      421 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
      481 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
      541 ttgcaaagaa atgaatatca gagagtgaga ggccttgaca ttgctagcgt ttaccgtcga
      601 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
      661 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
      721 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
      781 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
      841 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      901 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      961 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1021 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1081 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1141 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1201 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1261 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1321 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1381 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1441 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1501 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1561 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1621 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1681 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1741 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1801 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     1861 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     1921 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1981 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     2041 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     2101 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     2161 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     2221 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     2281 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     2341 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     2401 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     2461 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     2521 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     2581 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     2641 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     2701 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     2761 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcaacttgtt tattgcagct
     2821 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2881 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac
     2941 tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact
     3001 gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt
     3061 taaagaaatt gtatttgtta aatatgtact acaaacttag tagttggaag ggctaattca
     3121 ctcccaaaga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     3181 tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg
     3241 ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac
     3301 cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga
     3361 gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta
     3421 cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg
     3481 gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc
     3541 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     3601 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     3661 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     3721 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
     3781 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     3841 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     3901 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     3961 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     4021 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     4081 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     4141 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     4201 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg
     4261 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     4321 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     4381 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     4441 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     4501 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     4561 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     4621 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     4681 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     4741 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     4801 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     4861 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     4921 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     4981 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     5041 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt
     5101 cgattagtga acggatctcg acggtatcgc ctttaaaaga aaagggggga ttggggggta
     5161 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     5221 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
     5281 ttatcgatga ggccctttcg tcttcactcg aggtgcccgt cagtgggcag agcgcacatc
     5341 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
     5401 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     5461 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     5521 tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg
     5581 ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc
     5641 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
     5701 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
     5761 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
     5821 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
     5881 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
     5941 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
     6001 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
     6061 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
     6121 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
     6181 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     6241 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     6301 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     6361 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     6421 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     6481 gtgaggatcc gcggccgcgc cggctctaga tcgcgaacgc gtgaattcct cgagggcggc
     6541 cgctctagag tcgacgggcc gcggtaacaa ttgttaacta acttaagcta gcaacggttt
     6601 ccctctagcg ggatcaattc cgcccccccc ccctaacgtt actggccgaa gccgcttgga
     6661 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     6721 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     6781 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     6841 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     6901 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     6961 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     7021 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     7081 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     7141 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga taataccatg accgagtaca
     7201 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     7261 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     7321 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     7381 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     7441 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     7501 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     7561 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     7621 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     7681 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     7741 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccc gggcggggcg
     7801 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     7861 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     7921 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     7981 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     8041 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     8101 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     8161 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     8221 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     8281 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     8341 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     8401 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca
     8461 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt
     8521 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     8581 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat
     8641 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca
     8701 ccagggccag gggtcagata tccactgacc tttggatg
//
