LOCUS       V005060                 8664 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005060
VERSION     V005060
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8664)
  AUTHORS   De Groote P, Grootjans S, Lippens S, Eichperger C, Leurs K, Kahr I,
            Tanghe G, Bruggeman I, De Schamphelaire W, Urwyler C, Vandenabeele
            P, Haustraete J, Declercq W.
  TITLE     Generation of a new Gateway-compatible inducible lentiviral vector
            platform allowing easy derivation of co-transduced cells
  JOURNAL   BioTechniques 60 (5), 252-259 (2016)
   PUBMED   27177818
REFERENCE   2  (bases 1 to 8664)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-DEC-2015) BCCM/LMBP, Universiteit Gent,
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 8664)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8664)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "BioTechniques"; date: "2016"; volume: "60"; issue: "5"; pages:
            "252-259"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (22-DEC-2015) BCCM/LMBP, Universiteit Gent, Technologiepark 927,
            9052, BELGIUM"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..16
                     /label="HIV-1 cPPT"
                     /note="HIV-1 cPPT"
     LTR             18..251
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    323..457
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      484..619
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(640..658)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(668..684)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      826..1281
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1307..1411
                     /label="AmpR promoter"
     CDS             1412..2269
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      2443..3031
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    3319..3340
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3355..3385
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3393..3409
                     /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     3417..3433
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        3454..3472
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3500..3726
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3727..3907
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3954..4079
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    4572..4805
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             4990..5034
                     /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             5183..5224
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     enhancer        5313..5616
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5617..5820
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    5937..6061
                     /label="attR1"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        6086..6116
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             6170..6826
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     CDS             7171..7473
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(7517..7641)
                     /label="attR2"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             7685..7708
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     promoter        7753..8083
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        8131..8178
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             8197..8592
                     /label="BSD"
                     /note="blasticidin S deaminase"
ORIGIN
        1 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
       61 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
      121 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
      181 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
      241 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
      301 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
      361 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
      421 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
      481 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
      541 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
      601 ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg
      661 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
      721 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
      781 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
      841 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      901 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
      961 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1021 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1081 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1141 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1201 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     1261 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1321 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1381 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1441 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1501 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1561 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1621 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     1681 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1741 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1801 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     1861 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     1921 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     1981 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2041 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2101 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2161 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2221 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2281 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2341 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2401 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2461 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2521 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2581 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2641 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2701 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2761 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2821 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2881 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2941 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3001 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3061 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3121 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3181 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     3241 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     3301 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     3361 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     3421 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
     3481 gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta
     3541 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg
     3601 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt
     3661 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac
     3721 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     3781 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     3841 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     3901 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     3961 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     4021 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     4081 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     4141 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     4201 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     4261 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     4321 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     4381 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     4441 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     4501 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     4561 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     4621 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     4681 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     4741 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     4801 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     4861 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     4921 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     4981 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     5041 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     5101 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     5161 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
     5221 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
     5281 tctcgacggt atcgataagc ttgggagttc cgcgttacat aacttacggt aaatggcccg
     5341 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     5401 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     5461 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     5521 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     5581 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     5641 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     5701 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     5761 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     5821 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
     5881 agacaccgac tctagaggat ccactagtcc agtgtggtgg aattctgcag atatcaacaa
     5941 gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt
     6001 agattttgca taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat
     6061 ggcggccgca ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg
     6121 gattttgagt taggatccgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     6181 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     6241 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     6301 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     6361 ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat
     6421 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     6481 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     6541 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     6601 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     6661 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     6721 gatgccgctg gcgattcagg ttcatcatgc cgtctgtgat ggcttccatg tcggcagaat
     6781 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaaa gatctggatc
     6841 cggcttacta aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag
     6901 aatatatact gatatgtata cccgaagtat gtcaaaaaga ggtgtgctat gaagcagcgt
     6961 attacagtga cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata
     7021 tctccggtct ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct
     7081 ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct
     7141 cttttgctga cgagaacagg gactggtgaa atgcagttta aggtttacac ctataaaaga
     7201 gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga
     7261 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt
     7321 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
     7381 gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
     7441 aaaaacgcca ttaacctgat gttctgggga atataaatgt caggctccgt tatacacagc
     7501 cagtctgcag gtcgaccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg
     7561 ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc
     7621 agctttcttg tacaaagtgg ttgatatcca gcacagtggc ggccgctcga gtctagaggg
     7681 cccggactac aaggatgacg atgacaaaac gcgtaccggt tagtaatgag tttggaatta
     7741 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc aggcaggcag
     7801 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     7861 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     7921 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     7981 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     8041 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     8101 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
     8161 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa
     8221 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
     8281 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
     8341 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
     8401 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
     8461 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg
     8521 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat
     8581 gtgtgggagg gctaagcaca attcgagctc ggtaccttta agaccaatga cttacaaggc
     8641 agctgtagat cttagccact tttt
//
