LOCUS       V008022                 8369 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V008022
VERSION     V008022
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8369)
  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 8369)
  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 8369)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8369)
  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..8369
                     /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..1815
                     /label="VSV-G tag"
                     /note="epitope tag from vesicular stomatitis virus G
                     protein"
     promoter        1860..2190
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        2238..2285
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             2304..2699
                     /label="BSD"
                     /note="blasticidin S deaminase"
     misc_feature    2772..2787
                     /label="HIV-1 cPPT"
                     /note="HIV-1 cPPT"
     LTR             2789..3022
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3094..3228
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      3255..3390
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(3411..3429)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3439..3455)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      3597..4052
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4078..4182
                     /label="AmpR promoter"
     CDS             4183..5040
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5214..5802
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    6090..6111
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6126..6156
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    6164..6180
                     /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     6188..6204
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        6225..6243
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6271..6497
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6498..6678
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6725..6850
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7343..7576
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             7761..7805
                     /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             7954..7995
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     protein_bind    8093..8111
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8128..8146
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8164..8182
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8200..8218
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8235..8253
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8271..8289
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        8301..8339
                     /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 cgtacaccga catcgagatg
     1801 aacagactgg gcaagacgcg taccggttag taatgagttt ggaattaatt ctgtggaatg
     1861 tgtgtcagtt agggtgtgga aagtccccag gctccccagg caggcagaag tatgcaaagc
     1921 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     1981 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2041 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2101 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     2161 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     2221 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
     2281 acgacaaggt gaggaactaa accatggcca agcctttgtc tcaagaagaa tccaccctca
     2341 ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca
     2401 gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg
     2461 ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc
     2521 tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt
     2581 gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg
     2641 gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct
     2701 aagcacaatt cgagctcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     2761 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     2821 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     2881 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     2941 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     3001 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     3061 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     3121 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3181 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     3241 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3301 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3361 agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga
     3421 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     3481 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     3541 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     3601 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3661 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3721 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3781 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3841 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3901 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3961 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4021 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     4081 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4141 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4201 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4261 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4321 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4381 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4441 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4501 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4561 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4621 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4681 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4741 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4801 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4861 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4921 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4981 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5041 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5101 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5161 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5221 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5281 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5341 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5401 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5461 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5521 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5581 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5641 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5701 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5761 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5821 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5881 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     5941 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     6001 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     6061 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     6121 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     6181 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     6241 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
     6301 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
     6361 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
     6421 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
     6481 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6541 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6601 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6661 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
     6721 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     6781 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     6841 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     6901 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     6961 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     7021 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     7081 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     7141 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     7201 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     7261 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     7321 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     7381 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     7441 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     7501 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     7561 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     7621 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     7681 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     7741 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     7801 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     7861 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     7921 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg
     7981 acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat
     8041 tagtgaacgg atctcgacgg tatcgatggt ccgaggttct agacgagttt actccctatc
     8101 agtgatagag aacgatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgagtt
     8161 tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac
     8221 gtatgtcgag tttatcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag
     8281 tgatagagaa cgtatgtcga ggtaggcgtg tacggtggga ggcctatata agcagagctc
     8341 gtttagtgaa ccgtcagatc gcaccggtc
//
