LOCUS       V007540                 9452 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V007540
VERSION     V007540
KEYWORDS    pLenti CMV V5-LUC Blast (w567-1)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9452)
  AUTHORS   Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO,
            Campisi J, Yaswen P, Cooper PK, Kaufman PD.
  TITLE     A Versatile Viral System for Expression and Depletion of Proteins in
            Mammalian Cells
  JOURNAL   PLoS One. 2009 Aug 6;4(8):e6529.
   PUBMED   19657394
REFERENCE   2  (bases 1 to 9452)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9452)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2009-08-6"; volume: "4(8)"; pages: "e6529"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..130
                     /label="AmpR promoter"
     CDS             131..988
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1162..1750
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     1904..1921
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     protein_bind    2038..2059
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2074..2104
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2112..2128
                     /label="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)."
     primer_bind     2136..2152
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        2173..2191
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2219..2445
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2446..2626
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2673..2798
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    3291..3524
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             3709..3753
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             3902..3943
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    4020..4136
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        4159..4462
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4463..4666
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     4663..4687
                     /label="LNCX"
                     /note="Human CMV promoter, forward primer"
     protein_bind    4771..4795
                     /label="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4807..4848
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     CDS             4924..6573
                     /label="luciferase"
                     /note="firefly luciferase"
     protein_bind    complement(6583..6607)
                     /label="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    6638..7226
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     promoter        7282..7611
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        7659..7706
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             7725..8120
                     /label="BSD"
                     /note="blasticidin S deaminase"
     LTR             8210..8443
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8515..8649
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      8655..8790
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(8811..8829)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8839..8855)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      8997..9452
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
       61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2161 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
     2221 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     2281 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     2341 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     2401 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     2461 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     2521 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     2581 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     2641 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     2701 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     2761 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     2821 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     2881 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     2941 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     3001 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     3061 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     3121 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     3181 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     3241 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     3301 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     3361 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     3421 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     3481 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     3541 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     3601 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     3661 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     3721 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     3781 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     3841 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     3901 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     3961 gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact
     4021 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     4081 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaattcaa aattttatcg
     4141 ataagcttgg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     4201 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     4261 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     4321 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     4381 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     4441 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     4501 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     4561 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     4621 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
     4681 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgactcta
     4741 gtccagtgtg gtggaattct gcagatatca acaagtttgt acaaaaaagc aggctccacc
     4801 atggaaggta agcctatccc taaccctctc ctcggtctcg attctacggg atccatggga
     4861 accaattcag tccctaggct tttgcaaaaa gcttggcatt ccggtactgt tggtaaagcc
     4921 accatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc gctggaagat
     4981 ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca
     5041 attgctttta cagatgcaca tatcgaggtg gacatcactt acgctgagta cttcgaaatg
     5101 tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc
     5161 gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga
     5221 gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgggc
     5281 atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg
     5341 caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg attctaaaac ggattaccag
     5401 ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac
     5461 gattttgtgc cagagtcctt cgatagggac aagacaattg cactgatcat gaactcctct
     5521 ggatctactg gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg cgtgagattc
     5581 tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt
     5641 gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga
     5701 tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttctgaggag ccttcaggat
     5761 tacaagattc aaagtgcgct gctggtgcca accctattct ccttcttcgc caaaagcact
     5821 ctgattgaca aatacgattt atctaattta cacgaaattg cttctggtgg cgctcccctc
     5881 tctaaggaag tcggggaagc ggttgccaag aggttccatc tgccaggtat caggcaagga
     5941 tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg
     6001 ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg
     6061 aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc
     6121 ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat
     6181 tctggagaca tagcttactg ggacgaagac gaacacttct tcatcgttga ccgcctgaag
     6241 tctctgatta agtacaaagg ctatcaggtg gctcccgctg aattggaatc catcttgctc
     6301 caacacccca acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa
     6361 cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg
     6421 gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg
     6481 gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc
     6541 ataaaggcca agaagggcgg aaagatcgcc gtgtaattct agacccagct ttcttgtaca
     6601 aagtggttga tatccagcac agtggcggcc gctcgacaat caacctctgg attacaaaat
     6661 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     6721 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     6781 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     6841 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     6901 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     6961 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     7021 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     7081 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     7141 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     7201 gatctccctt tgggccgcct ccccgcctgg aattctgcag atatccggtt agtaatgagt
     7261 ttggaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     7321 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     7381 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     7441 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     7501 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     7561 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     7621 ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat
     7681 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc aagcctttgt
     7741 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
     7801 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
     7861 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
     7921 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
     7981 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
     8041 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
     8101 ctggttatgt gtgggagggc taagcacaat tcgagctcgg tacctttaag accaatgact
     8161 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     8221 gctcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     8281 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     8341 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     8401 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     8461 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     8521 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     8581 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     8641 gtctggctct agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     8701 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     8761 agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga
     8821 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     8881 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     8941 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     9001 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     9061 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     9121 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     9181 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     9241 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     9301 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     9361 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     9421 gaattttaac aaaatattaa cgcttacaat tt
//
