LOCUS       V008026                 9098 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V008026
VERSION     V008026
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9098)
  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 9098)
  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 9098)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9098)
  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..9098
                     /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             1269..1571
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(1615..1739)
                     /label="attR2"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1771..2487
                     /label="EGFP"
                     /note="enhanced GFP"
     CDS             2506..2547
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label="V5 tag"
                     /translation="GKPIPNPLLGLDST"
     promoter        2589..2919
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        2967..3014
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             3033..3428
                     /label="BSD"
                     /note="blasticidin S deaminase"
     misc_feature    3501..3516
                     /label="HIV-1 cPPT"
                     /note="HIV-1 cPPT"
     LTR             3518..3751
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3823..3957
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      3984..4119
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(4140..4158)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4168..4184)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      4326..4781
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4807..4911
                     /label="AmpR promoter"
     CDS             4912..5769
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5943..6531
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    6819..6840
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6855..6885
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    6893..6909
                     /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     6917..6933
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        6954..6972
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7000..7226
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7227..7407
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7454..7579
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    8072..8305
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             8490..8534
                     /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             8683..8724
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     protein_bind    8822..8840
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8857..8875
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8893..8911
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8929..8947
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8964..8982
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9000..9018
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        9030..9068
                     /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 ggcgtaaaga tctggatccg gcttactaaa agccagataa
      961 cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga tatgtatacc
     1021 cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca gttgacagcg
     1081 acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg taagcacaac
     1141 catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg
     1201 gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg agaacaggga
     1261 ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg
     1321 tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc cccctggcca
     1381 gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg
     1441 atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg
     1501 aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt
     1561 tctggggaat ataaatgtca ggctccgtta tacacagcca gtctgcaggt cgaccatagt
     1621 gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt
     1681 aatatattga tatttatatc attttacgtt tctcgttcag ctttcttgta caaagtggtt
     1741 gatatccagc acagtggcgg ccgctcgagt atggtgagca agggcgagga gctgttcacc
     1801 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1861 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1921 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     1981 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     2041 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     2101 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2161 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2221 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     2281 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2341 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     2401 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2461 actctcggca tggacgagct gtacaaggag ggcccgcggt tcgaaggtaa gcctatccct
     2521 aaccctctcc tcggtctcga ttctacgcgt accggttagt aatgagtttg gaattaattc
     2581 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccaggc aggcagaagt
     2641 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2701 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2761 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2821 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2881 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2941 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     3001 tagtataata cgacaaggtg aggaactaaa ccatggccaa gcctttgtct caagaagaat
     3061 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     3121 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     3181 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     3241 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     3301 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     3361 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     3421 gggagggcta agcacaattc gagctcggta cctttaagac caatgactta caaggcagct
     3481 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     3541 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     3601 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     3661 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     3721 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     3781 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     3841 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3901 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     3961 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4021 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4081 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc
     4141 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     4201 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     4261 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     4321 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4381 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4441 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4501 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4561 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4621 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4681 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4741 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     4801 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4861 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4921 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4981 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5041 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5101 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5161 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5221 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5281 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5341 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5401 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5461 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5521 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5581 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5641 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5701 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5761 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5821 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5881 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5941 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6001 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6061 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6121 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6181 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6241 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6301 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6361 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6421 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6481 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6541 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6601 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     6661 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     6721 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     6781 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     6841 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     6901 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
     6961 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
     7021 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
     7081 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
     7141 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
     7201 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
     7261 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     7321 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     7381 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     7441 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     7501 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     7561 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     7621 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     7681 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     7741 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     7801 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     7861 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     7921 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     7981 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     8041 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     8101 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     8161 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     8221 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     8281 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     8341 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     8401 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     8461 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     8521 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     8581 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     8641 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     8701 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     8761 ccattcgatt agtgaacgga tctcgacggt atcgatggtc cgaggttcta gacgagttta
     8821 ctccctatca gtgatagaga acgatgtcga gtttactccc tatcagtgat agagaacgta
     8881 tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
     8941 atagagaacg tatgtcgagt ttatccctat cagtgataga gaacgtatgt cgagtttact
     9001 ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt acggtgggag gcctatataa
     9061 gcagagctcg tttagtgaac cgtcagatcg caccggtc
//
