LOCUS       V015051                10203 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V015051
VERSION     V015051
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10203)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    complement(216..350)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        398..502
                     /label="AmpR promoter"
     CDS             503..1360
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1534..2119
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2407..2428
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2443..2473
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     LTR             complement(2650..3283)
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     misc_feature    complement(3491..4079)
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     CDS             complement(4096..4887)
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase from Tn5"
     promoter        complement(4896..5225)
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             complement(5239..5982)
                     /label="Tet-On(R) 3G"
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
     promoter        complement(6001..6511)
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     promoter        6528..6895
                     /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"
     CDS             6907..7623
                     /label="EGFP"
                     /note="enhanced GFP"
     CDS             7684..7707
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     misc_feature    complement(8060..8177)
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     CDS             complement(8249..8290)
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     CDS             complement(8439..8483)
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     misc_feature    complement(8668..8901)
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     misc_feature    complement(9398..9523)
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     LTR             complement(9570..10203)
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 actactaagt ttgtagtaca tatttaacaa atacaatttc tttaaaaact actaagtttg
       61 tagtacatat ttaacaaata caatttcttt aaaatgaaaa taattcagag gaatcacagg
      121 tttagagtaa atgaaaccac taggtaattg gcagtggtaa tagggtatgg ggtgggaagt
      181 ttgggatgat tttggttagc ttgagttatc cagttgatcc agacatgata agatacattg
      241 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      301 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt gatctcccga
      361 tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      421 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      481 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      541 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      601 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      661 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      721 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
      781 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
      841 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      901 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      961 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1021 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1081 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1141 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1201 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1261 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1321 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1381 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1441 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1501 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1561 ctgctgcttg caaacaaaaa aaccgctacc agcggtggtt tgtttgccgg atcaagagct
     1621 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1681 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1741 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1801 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1861 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1921 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1981 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2041 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2101 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2161 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2221 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2281 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2341 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     2401 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     2461 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     2521 ccatgattac gccaagctct agctagaggt cgacggtaaa acgctagcaa tgtcaaggcc
     2581 tctcactctc tgatattcat ttctttgcaa gttataaata ctgaataata agatgacatg
     2641 aactactact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     2701 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     2761 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     2821 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
     2881 tccccagtcc cgcccaggcc acgcctccct ggaaagtccc cagcggaaag tcccttgtag
     2941 caagctcgat atcagcagtt cttgaagtac tccggatgca gctctcgggc cacgtgatga
     3001 aatgctaggc ggctgtcaaa cctccactct aacacttctc tctccgggtc atccatccca
     3061 tgcaggctca cagggtgtaa caagctggtg ttctctcctt tattggcctc ttctacctta
     3121 tctggctcaa ctggtactag cttgtagcac catccaaagg tcagtggata tctgacccct
     3181 ggccctggtg tgtagttctg ctaatcaggg aagtagcctt gtgtgtggta gatccacaga
     3241 tcaaggatat cttgtcttcg ttgggagtga attagccctt ccagtcccct cttttctttt
     3301 aaaaagtggc taagatctac agctgccttg taagtcattg gtcttaaagg tacctgaggt
     3361 gtgactggaa aaacccacct cctcctcctc ttgtgcttct agccaggcac aatcagcatt
     3421 ggtagctgct gtattgctac ttgtgattgc tccatgtttt tctaggtctc gactgcagaa
     3481 ttaattccag gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga
     3541 agacgcggaa gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg
     3601 agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacacaggcg
     3661 agcagccatg gaaaggacgt cagcttcccc gacaacacca cggaattgtc agtgcccaac
     3721 agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata
     3781 gggaggggga aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca
     3841 gtgggggttg cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac
     3901 aactcctcat aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata
     3961 cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga
     4021 gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgattg
     4081 ttccagacgc gttcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     4141 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     4201 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc
     4261 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     4321 gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag
     4381 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
     4441 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
     4501 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
     4561 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
     4621 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
     4681 ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt
     4741 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
     4801 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
     4861 tgcgtgcaat ccatcttgtt cagccatggt cgcgttttgc aaaagcctag gcctccaaaa
     4921 aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa
     4981 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     5041 gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
     5101 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga
     5161 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
     5221 cacacattcg cgcgtttacc cggggagcat gtcaaggtca aaatcgtcaa gagcgtcagc
     5281 aggcagcata tcaaggtcaa agtcgtcaag ggcatcggct gggagcatgt ctaagtcaaa
     5341 atcgtcaagg gcgtcggtcg gcccgccgct ttcgcacttt agctgtttct ccaggccaca
     5401 tatgattagt tccaggccga aaaggaaggc aggttcggct ccctgccggt cgaacagctc
     5461 aattgcttgt ttcagaagtg ggggcataga atcggtggta ggtgtctctc tttcctcttt
     5521 tgctacttga tgctcctgtt cctccaatac gcagcccagt gtaaagtggc ccacggcgga
     5581 cagagcgtac agtgcgttct ccagggagaa gccttgctga cacaggaacg cgagctgatt
     5641 ttccagggtt tcgtactgtt tctctgttgg gcgggtgccg agatgcactt tagccccgtc
     5701 gcgatgtgag aggagagcac agcggtatga cttggcgttg ttccgcagaa agtcttgcca
     5761 tgactcgcct tccagggggc aggagtgggt atgatgcctg tccagcatct cgattggcag
     5821 ggcatcgagc agggcccgct tgttcttcac gtgccagtac agggtaggct gctcaactcc
     5881 cagcttttga gcgagtttcc ttgtcgtcag gccttcgata ccgactccat tgagtaattc
     5941 cagagcagag tttatgactt tgctcttgtc cagtctagac atggtaattc gatgatcccc
     6001 ctggggagag aggtcggtga ttcggtcaac gagggagccg actgccgacg tgcgctccgg
     6061 aggcttgcag aatgcggaac accgcgcggg caggaacagg gcccacacta ccgccccaca
     6121 ccccgcctcc cgcaccgccc cttcccggcc gctgctctcg gcgcgccctg ctgagcagcc
     6181 gctattggcc acagcccatc gcggtcggcg cgctgccatt gctccctggc gctgtccgtc
     6241 tgcgagggta ctagtgagac gtgcggcttc cgtttgtcac gtccggcacg ccgcgaaccg
     6301 caaggaacct tcccgactta ggggcggagc aggaagcgtc gccggggggc ccacaagggt
     6361 agcggcgaag atccgggtga cgctgcgaac ggacgtgaag aatgtgcgag acccagggtc
     6421 ggcgccgctg cgtttcccgg aaccacgccc agagcagccg cgtccctgcg caaacccagg
     6481 gctgccttgg aaaaggcgca accccaaccc cgtggaatta tcacctcgag tttactccct
     6541 atcagtgata gagaacgtat gaagagttta ctccctatca gtgatagaga acgtatgcag
     6601 actttactcc ctatcagtga tagagaacgt ataaggagtt tactccctat cagtgataga
     6661 gaacgtatga ccagtttact ccctatcagt gatagagaac gtatctacag tttactccct
     6721 atcagtgata gagaacgtat atccagttta ctccctatca gtgatagaga acgtataagc
     6781 tttgcttatg taaaccaggg cgcctataaa agagtgctga ttttttgagt aaacttcaat
     6841 tccacaacac ttttgtctta taccaacttt ccgtaccact tcctaccctc gtaaatcagt
     6901 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     6961 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     7021 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     7081 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     7141 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     7201 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     7261 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     7321 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     7381 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     7441 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     7501 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     7561 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     7621 aaggaattca ccggtggatc cgactacaaa gaccatgacg gtgattataa agatcatgac
     7681 atcgactaca aggatgacga tgacaagtag accacctccc ctgcgagcta agctggacag
     7741 ccaatgacgg gtaagagagt gacatttttc actaacctaa gacaggaggg ccgtcagagc
     7801 tactgcctaa tccaaagacg ggtaaaagtg ataaaaatgt atcactccaa cctaagacag
     7861 gcgcagcttc cgagggattt gagatccaga catgataaga tacattgatg agtttggaca
     7921 aaccaaaact agaatgcagt gaaaaaaatg ccttatttgt gaaatttgtg atgctattgc
     7981 cttatttgta accattataa gctgcaataa acaagttaag tcgataaact ggatctctgc
     8041 tgtccctgta ataaacccga aaattttgaa tttttgtaat ttgtttttgt agttctttag
     8101 tttgtatgtc tgttgctatt atgtctacta ttctttcccc tgcactgtac cccccaatcc
     8161 ccccttttct tttaaaggcg ataccgtcga gatccgttca ctaatcgaat ggatctgtct
     8221 ctgtctctct ctccaccttc ttcttctatt ccttcgggcc tgtcgggtcc cctcggggtt
     8281 gggaggtggg tctgaaacga taatggtgaa tatccctgcc taactctatt cactatagaa
     8341 agtacagcaa aaactattct taaacctacc aagcctccta ctatcattat gaataatttt
     8401 atataccaca gccaatttgt tatgttaaac caattccaca aacttgccca tttatctaat
     8461 tccaataatt cttgttcatt cttttcttgc tggttttgcg attcttcaat taaggagtgt
     8521 attaagcttg tgtaattgtt aatttctctg tcccactcca tccaggtcgt gtgattccaa
     8581 atctgttcca gagatttatt actccaacta gcattccaag gcacagcagt ggtgcaaatg
     8641 agttttccag agcaacccca aatccccagg agctgttgat cctttaggta tctttccaca
     8701 gccaggattc ttgcctggag ctgcttgatg ccccagactg tgagttgcaa cagatgctgt
     8761 tgcgcctcaa tagccctcag caaattgttc tgctgctgca ctataccaga caataattgt
     8821 ctggcctgta ccgtcagcgt cattgacgct gcgcccatag tgcttcctgc tgctcccaag
     8881 aacccaagga acaaagctcc tattcccact gctctttttt ctctctgcac cactcttctc
     8941 tttgccttgg tgggtgctac tcctaatggt tcaattttta ctactttata tttatataat
     9001 tcacttctcc aattgtccct catatctcct cctccaggtc tgaagatcag cggccggccg
     9061 cttgctgtgc ggtggtctta cttttgtttt gctcttcctc tatcttgtct aaagcttcct
     9121 tggtgtcttt tatctctatc ctttgatgca cacaatagag ggttgctact gtattatata
     9181 atgatctaag ttcttctgat cctgtctgaa gggatggttg tagctgtccc agtatttgtc
     9241 tacagccttc tgatgtttct aacaggccag gattaactgc gaatcgttct agctccctgc
     9301 ttgcccatac tatatgtttt aatttatatt ttttctttcc ccctggcctt aaccgaattt
     9361 tttcccatcg cgatctaatt ctcccccgct taatactgac gctctcgcac ccatctctct
     9421 ccttctagcc tccgctagtc aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc
     9481 ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga gagctcctct ggtttccctt
     9541 tcgctttcaa gtccctgttc gggcgccact gctagagatt ttccacactg actaaaaggg
     9601 tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac
     9661 tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag agagctccca
     9721 ggctcagatc tggtctaacc agagagaccc agtacaggca aaaagcagct gcttatatgc
     9781 aggatctgag ggctcgccac tccccagtcc cgcccaggcc acgcctccct ggaaagtccc
     9841 cagcggaaag tcccttgtag caagctcgat atcagcagtt cttgaagtac tccggatgca
     9901 gctctcgggc cacgtgatga aatgctaggc ggctgtcaaa cctccactct aacacttctc
     9961 tctccgggtc atccatccca tgcaggctca cagggtgtaa caagctggtg ttctctcctt
    10021 tattggcctc ttctacctta tctggctcaa ctggtactag cttgtagcac catccaaagg
    10081 tcagtggata tctgacccct ggccctggtg tgtagttctg ctaatcaggg aagtagcctt
    10141 gtgtgtggta gatccacaga tcaaggatat cttgtcttct ttgggagtga attagccctt
    10201 cca
//
