LOCUS       V010419                 9227 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010419
VERSION     V010419
KEYWORDS    pLVX-TetOne-Puro.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9227)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9227)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1303..1536
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1721..1765
                     /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             1914..1955
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2027..2144
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     polyA_signal    2187..2321
                     /label="SV40 poly(A) signal"
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     misc_feature    2496..2527
                     /label="MCS"
                     /note="MCS"
                     /note="multiple cloning site"
     promoter        complement(2528..2895)
                     /label="TRE3GS promoter"
                     /note="3rd-generation Tet-responsive promoter that can be
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription
                     factors"
     promoter        2912..3422
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3441..4184
                     /label="Tet-On(R) 3G"
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
     promoter        4198..4527
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             4536..5132
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    5149..5737
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5945..6578
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6707..6723)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6731..6747)
                     /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)."
     promoter        complement(6755..6785)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6800..6821)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7109..7694)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7868..8725)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8726..8830)
                     /label="AmpR promoter"
     polyA_signal    8878..9012
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga actacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgacttaact tgtttattgc agcttataat ggttacaaat
     2221 aaggcaatag catcacaaat ttcacaaata aggcattttt ttcactgcat tctagttttg
     2281 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctcaaatccc tcggaagctg
     2341 cgcctgtctt aggttggagt gatacatttt tatcactttt acccgtcttt ggattaggca
     2401 gtagctctga cggccctcct gtcttaggtt agtgaaaaat gtcactctct tacccgtcat
     2461 tggctgtcca gcttagctcg caggggaggt ggtctggatc cgccggcacc ggtgtatacg
     2521 ggaattcttt acgagggtag gaagtggtac ggaaagttgg tataagacaa aagtgttgtg
     2581 gaattgaagt ttactcaaaa aatcagcact cttttatagg cgccctggtt tacataagca
     2641 aagcttatac gttctctatc actgataggg agtaaactgg atatacgttc tctatcactg
     2701 atagggagta aactgtagat acgttctcta tcactgatag ggagtaaact ggtcatacgt
     2761 tctctatcac tgatagggag taaactcctt atacgttctc tatcactgat agggagtaaa
     2821 gtctgcatac gttctctatc actgataggg agtaaactct tcatacgttc tctatcactg
     2881 atagggagta aactcgaggt gataattcca cggggttggg gttgcgcctt ttccaaggca
     2941 gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg
     3001 ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg
     3061 ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct
     3121 tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg
     3181 cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata
     3241 gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg
     3301 ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc
     3361 ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc
     3421 agggggatca tcgaattacc atgtctagac tggacaagag caaagtcata aactctgctc
     3481 tggaattact caatggagtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc
     3541 tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg
     3601 ccctgccaat cgagatgctg gacaggcatc atacccactc ctgccccctg gaaggcgagt
     3661 catggcaaga ctttctgcgg aacaacgcca agtcataccg ctgtgctctc ctctcacatc
     3721 gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg
     3781 aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc
     3841 tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag
     3901 caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg aaacaagcaa
     3961 ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca
     4021 tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccgaccgac gcccttgacg
     4081 attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc
     4141 ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaaacg cgcgaatgtg
     4201 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4261 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4321 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4381 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4441 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4501 ttttttggag gcctaggctt ttgcaaaacg cgaccatgac cgagtacaag cccacggtgc
     4561 gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg
     4621 actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc
     4681 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4741 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4801 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4861 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4921 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4981 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     5041 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     5101 gcacctggtg catgacccgc aagcccggtg cctgaacgcg tctggaacaa tcaacctctg
     5161 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     5221 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     5281 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     5341 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     5401 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     5461 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     5521 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     5581 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     5641 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     5701 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc tgcagtcgag
     5761 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
     5821 cctggctaga agcacaagag gaggaggagg tgggtttttc cagtcacacc tcaggtacct
     5881 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg
     5941 ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac
     6001 cacacacaag gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat
     6061 ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag
     6121 gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac
     6181 ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc
     6241 cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt
     6301 tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc
     6361 agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat
     6421 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     6481 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     6541 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat
     6601 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg
     6661 ctagcgtttt accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6721 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6781 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6841 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6901 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6961 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7021 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7081 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7141 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7201 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7261 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7321 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7381 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7441 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7501 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7561 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7621 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7741 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7801 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7861 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7921 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7981 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     8041 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     8101 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     8161 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     8221 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8281 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8341 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8401 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8461 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8521 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8581 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8641 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8701 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8761 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8821 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac
     8881 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     8941 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     9001 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
     9061 taccctatta ccactgccaa ttacctagtg gtttcattta ctctaaacct gtgattcctc
     9121 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
     9181 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//
