LOCUS       V008025                 8363 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V008025
VERSION     V008025
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8363)
  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 8363)
  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 8363)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8363)
  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..8363
                     /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             1783..1809
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     promoter        1854..2184
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        2232..2279
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             2298..2693
                     /label="BSD"
                     /note="blasticidin S deaminase"
     misc_feature    2766..2781
                     /label="HIV-1 cPPT"
                     /note="HIV-1 cPPT"
     LTR             2783..3016
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3088..3222
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      3249..3384
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(3405..3423)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3433..3449)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      3591..4046
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4072..4176
                     /label="AmpR promoter"
     CDS             4177..5034
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5208..5796
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    6084..6105
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6120..6150
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    6158..6174
                     /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     6182..6198
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        6219..6237
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6265..6491
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6492..6672
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6719..6844
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7337..7570
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             7755..7799
                     /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             7948..7989
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     protein_bind    8087..8105
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8122..8140
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8158..8176
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8194..8212
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8229..8247
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8265..8283
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        8295..8333
                     /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 agaacagggg
     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 ggctccctta tacacagcca gtctgcaggt cgaccatagt
     1621 gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt
     1681 aatatattga tatttatatc attttacgtt tctcgttcag ctttcttgta caaagtggtt
     1741 gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtaccccta cgacgtgccc
     1801 gactacgcca cgcgtaccgg ttagtaatga gtttggaatt aattctgtgg aatgtgtgtc
     1861 agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat
     1921 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     1981 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2041 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2101 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     2161 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     2221 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     2281 aggtgaggaa ctaaaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
     2341 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
     2401 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
     2461 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
     2521 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
     2581 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
     2641 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagcac
     2701 aattcgagct cggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     2761 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     2821 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     2881 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     2941 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     3001 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     3061 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     3121 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3181 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     3241 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3301 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3361 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     3421 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     3481 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     3541 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     3601 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3661 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3721 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3781 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3841 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3901 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3961 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4021 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     4081 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4141 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4201 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4261 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4321 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4381 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4441 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4501 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4561 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4621 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4681 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4741 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4801 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4861 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4921 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4981 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5041 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5101 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5161 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5221 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5281 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5341 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5401 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5461 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5521 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5581 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5641 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5701 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5761 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5821 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5881 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5941 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     6001 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     6061 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     6121 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     6181 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     6241 caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca
     6301 tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat
     6361 tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat
     6421 ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc
     6481 gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6541 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6601 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6661 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
     6721 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
     6781 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
     6841 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
     6901 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
     6961 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
     7021 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
     7081 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
     7141 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac
     7201 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     7261 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     7321 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     7381 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     7441 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     7501 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     7561 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     7621 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     7681 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     7741 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     7801 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     7861 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     7921 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     7981 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     8041 acggatctcg acggtatcga tggtccgagg ttctagacga gtttactccc tatcagtgat
     8101 agagaacgat gtcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc
     8161 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
     8221 cgagtttatc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag
     8281 agaacgtatg tcgaggtagg cgtgtacggt gggaggccta tataagcaga gctcgtttag
     8341 tgaaccgtca gatcgcaccg gtc
//
