LOCUS       V013507                 9399 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013507
VERSION     V013507
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9399)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9399
                     /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"
     misc_RNA        complement(2194..2269)
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     promoter        complement(2294..2534)
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     enhancer        2551..2854
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2855..3058
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             3169..3912
                     /label="Tet-On(R) 3G"
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
     misc_feature    3924..4509
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             4510..5301
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase from Tn5"
     misc_feature    5318..5906
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6114..6747
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6876..6892)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6900..6916)
                     /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(6924..6954)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6969..6990)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7278..7866)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8040..8897)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8898..9002)
                     /label="AmpR promoter"
     polyA_signal    9050..9184
                     /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 cgataaaaaa aaagcaccga ctcggtgcca ctttttcaag
     2221 ttgataacgg actagcctta ttttaacttg ctatttctag ctctaaaact gagacgtccg
     2281 tctccggtgt ttcgtccttt ccacaagata tataaagcca agaaatcgaa atactttcaa
     2341 gttacggtaa gcatatgata gtccatttta aaacataatt ttaaaactgc aaactaccca
     2401 agaaattatt actttctacg tcacgtattt tgtactaata tctttgtgtt tacagtcaaa
     2461 ttaattccaa ttatctctct aacagccttg tatcgtatat gcaaatatga aggaatcatg
     2521 ggaaataggc cctcaagctt gggagttccg cgttacataa cttacggtaa atggcccgcc
     2581 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     2641 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     2701 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     2761 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     2821 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     2881 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     2941 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     3001 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     3061 tttagtgaac cgtcagatcg cctggagacc atccacgctg ttttgacctc catagaagac
     3121 accgactcta ctagaggatc tatttccggt gaattcctcg aggccaccat gtctagactg
     3181 gacaagagca aagtcataaa ctctgctctg gaattactca atggagtcgg tatcgaaggc
     3241 ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct gtactggcac
     3301 gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg agatgctgga caggcatcat
     3361 acccactcct gccccctgga aggcgagtca tggcaagact ttctgcggaa caacgccaag
     3421 tcataccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct cggcacccgc
     3481 ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc
     3541 ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac actgggctgc
     3601 gtattggagg aacaggagca tcaagtagca aaagaggaaa gagagacacc taccaccgat
     3661 tctatgcccc cacttctgaa acaagcaatt gagctgttcg accggcaggg agccgaacct
     3721 gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct aaagtgcgaa
     3781 agcggcgggc cgaccgacgc ccttgacgat tttgacttag acatgctccc agccgatgcc
     3841 cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt tgaccttgac
     3901 atgctccccg ggtaaggatc ccgcccctct ccctcccccc cccctaacgt tactggccga
     3961 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     4021 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     4081 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     4141 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     4201 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     4261 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     4321 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     4381 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaac
     4441 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt
     4501 gccacaacca tggctgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4561 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4621 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4681 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4741 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4801 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4861 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4921 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4981 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5041 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5101 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5161 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5221 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5281 cgccttcttg acgagttctt ctgaacgcgt ctggaacaat caacctctgg attacaaaat
     5341 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5401 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5461 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5521 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5581 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5641 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5701 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     5761 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5821 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5881 gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa
     5941 catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa
     6001 gcacaagagg aggaggaggt gggtttttcc agtcacacct caggtacctt taagaccaat
     6061 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg
     6121 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
     6181 ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt
     6241 tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg
     6301 agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     6361 agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca
     6421 tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga
     6481 ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat
     6541 ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6601 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6661 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6721 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6781 tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttta
     6841 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6901 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6961 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     7021 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7081 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7141 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7201 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7261 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7321 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7381 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7441 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7501 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7561 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7621 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7681 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7741 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7801 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7861 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7921 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7981 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     8041 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     8101 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     8161 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     8221 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8281 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8341 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8401 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8461 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8521 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8581 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8641 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8701 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8761 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8821 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8881 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8941 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     9001 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat
     9061 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     9121 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     9181 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat
     9241 taccactgcc aattacctag tggtttcatt tactctaaac ctgtgattcc tctgaattat
     9301 tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta gtttttaaag
     9361 aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
