LOCUS       V008023                 8382 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V008023
VERSION     V008023
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8382)
  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 8382)
  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 8382)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8382)
  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..8382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    35..159
                     /label="attR1"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        184..214
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             268..924
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     CDS             1267..1569
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(1613..1737)
                     /label="attR2"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1790..1831
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     promoter        1873..2203
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        2251..2298
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             2317..2712
                     /label="BSD"
                     /note="blasticidin S deaminase"
     misc_feature    2785..2800
                     /label="HIV-1 cPPT"
                     /note="HIV-1 cPPT"
     LTR             2802..3035
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3107..3241
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      3268..3403
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(3424..3442)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3452..3468)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      3610..4065
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4091..4195
                     /label="AmpR promoter"
     CDS             4196..5053
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5227..5815
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    6103..6124
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6139..6169
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    6177..6193
                     /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     6201..6217
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        6238..6256
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6284..6510
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6511..6691
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6738..6863
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7356..7589
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             7774..7818
                     /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             7967..8008
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     protein_bind    8106..8124
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8141..8159
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8177..8195
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8213..8231
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8248..8266
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8284..8302
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        8314..8352
                     /label="minimal CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
ORIGIN
        1 actagtccag tgtggtggaa ttctgcagat atcaacaagt ttgtacaaaa aagctgaacg
       61 agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata aaaaacagac
      121 tacataatac tgtaaaacac aacatatcca gtcactatgg cggccgcatt aggcacccca
      181 ggctttacac tttatgcttc cggctcgtat aatgtgtgga ttttgagtta ggatccggcg
      241 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
      301 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
      361 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
      421 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
      481 gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
      541 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
      601 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
      661 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
      721 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
      781 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
      841 catcatgccg tctgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
      901 tgcgatgagt ggcagggcgg ggcgtagatc tggatccggc ttactaaaag ccagataaca
      961 gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg
     1021 aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac
     1081 agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca
     1141 tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga
     1201 tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct
     1261 ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
     1321 gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt
     1381 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
     1441 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
     1501 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
     1561 tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga
     1621 ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa
     1681 tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggttga
     1741 tatccagcac agtggcggcc gctcgagtct agagggcccg cggttcgaag gtaagcctat
     1801 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt tagtaatgag tttggaatta
     1861 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc aggcaggcag
     1921 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     1981 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2041 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2101 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     2161 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     2221 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
     2281 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa
     2341 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
     2401 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
     2461 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
     2521 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
     2581 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg
     2641 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat
     2701 gtgtgggagg gctaagcaca attcgagctc ggtaccttta agaccaatga cttacaaggc
     2761 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     2821 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     2881 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     2941 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     3001 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     3061 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     3121 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3181 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     3241 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3301 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3361 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
     3421 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     3481 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     3541 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     3601 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3661 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3721 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3781 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3841 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3901 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3961 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4021 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     4081 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4141 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4201 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4261 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4321 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4381 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4441 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4501 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4561 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4621 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4681 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4741 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4801 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4861 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4921 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4981 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5041 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5101 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5161 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5221 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5281 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5341 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5401 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5461 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5521 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5581 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5641 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5701 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5761 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5821 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5881 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5941 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     6001 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     6061 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     6121 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     6181 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     6241 taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata
     6301 ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa
     6361 aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg
     6421 caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt
     6481 gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc
     6541 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6601 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6661 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     6721 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     6781 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     6841 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     6901 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     6961 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     7021 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     7081 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     7141 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     7201 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     7261 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     7321 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     7381 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     7441 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     7501 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     7561 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     7621 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     7681 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     7741 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     7801 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     7861 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     7921 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     7981 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
     8041 agatccattc gattagtgaa cggatctcga cggtatcgat ggtccgaggt tctagacgag
     8101 tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag tgatagagaa
     8161 cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc
     8221 agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt atgtcgagtt
     8281 tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg ggaggcctat
     8341 ataagcagag ctcgtttagt gaaccgtcag atcgcaccgg tc
//
