LOCUS       V017224                 8648 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V017224
VERSION     V017224
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8648)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        626..644
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        657..958
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        959..1161
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             1176..1356
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1403..1528
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    2027..2260
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             2445..2489
                     /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             2638..2679
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2787..2904
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2979..3219
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     promoter        3354..3853
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             3893..4489
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     CDS             4568..5260
                     /label="ZsGreen1"
                     /note="Zoanthus green fluorescent protein"
     misc_feature    5271..5858
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    6210..6434
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(6460..6478)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6499..6515)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6523..6539)
                     /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(6547..6577)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6592..6613)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6901..7489)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7663..8520)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8521..8625)
                     /label="AmpR promoter"
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgactagtac
      661 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      721 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      781 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      841 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      901 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      961 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     1021 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     1081 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     1141 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
     1201 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     1261 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     1321 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     1381 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     1441 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     1501 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     1561 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     1621 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     1681 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     1741 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     1801 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     1861 gcaagcggcc ggccgcgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     1921 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1981 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     2041 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     2101 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     2161 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     2221 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     2281 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     2341 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     2401 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     2461 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     2521 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2581 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2641 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     2701 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     2761 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2821 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2881 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     2941 ttagtaccgg gccctacgcg tccaaggtcg ggcaggaaga gggcctattt cccatgattc
     3001 cttcatattt gcatatacga tacaaggctg ttagagagat aattagaatt aatttgactg
     3061 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     3121 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     3181 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccggg tcttcgagaa
     3241 gacacgttta agagctatgc tggaaacagc atagcaagtt taaataaggc tagtccgtta
     3301 tcaacttgaa aaagtggcac cgagtcggtg ctttttttgg atccaattct accgggtagg
     3361 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg
     3421 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc
     3481 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt
     3541 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac
     3601 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag
     3661 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc
     3721 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg
     3781 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc
     3841 tccgggcctt tcgacctgca tccatctaga tctcgagcag ctgaagctta ccatgaccga
     3901 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     3961 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     4021 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     4081 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     4141 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     4201 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     4261 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     4321 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     4381 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     4441 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgccg gcggcgggtc
     4501 cggaggagag ggcagaggaa gtctcctaac atgcggtgac gtggaggaga atcctggccc
     4561 agaattcatg gcccagtcca agcacggcct gaccaaggag atgaccatga agtaccgcat
     4621 ggagggctgc gtggacggcc acaagttcgt gatcaccggc gagggcatcg gctacccctt
     4681 caagggcaag caggccatca acctgtgcgt ggtggagggc ggccccttgc ccttcgccga
     4741 ggacatcttg tccgccgcct tcatgtacgg caaccgcgtg ttcaccgagt acccccagga
     4801 catcgtcgac tacttcaaga actcctgccc cgccggctac acctgggacc gctccttcct
     4861 gttcgaggac ggcgccgtgt gcatctgcaa cgccgacatc accgtgagcg tggaggagaa
     4921 ctgcatgtac cacgagtcca agttctacgg cgtgaacttc cccgccgacg gccccgtgat
     4981 gaagaagatg accgacaact gggagccctc ctgcgagaag atcatccccg tgcccaagca
     5041 gggcatcttg aagggcgacg tgagcatgta cctgctgctg aaggacggtg gccgcttgcg
     5101 ctgccagttc gacaccgtgt acaaggccaa gtccgtgccc cgcaagatgc ccgactggca
     5161 cttcatccag cacaagctga cccgcgagga ccgcagcgac gccaagaacc agaagtggca
     5221 cctgaccgag cacgccatcg cctccggctc cgccttgccc taagcggccg ctcaacctct
     5281 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     5341 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     5401 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     5461 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     5521 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     5581 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     5641 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     5701 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     5761 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtctac gccttcgccc
     5821 tcagacgagt cggatctccc tttgggccgc ctccccgcag gtacctttaa gaccaatgac
     5881 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     5941 aattcactcc caaagaagtc aagatctgct ttttgcctgt actgggtctc tctggttaga
     6001 ccagagtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6061 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6121 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6181 agcagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     6241 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     6301 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     6361 ggtggggcag gacagcaagg gggaggattg ggaagtcaat agcaggcatg ctggggatgc
     6421 ggtgggctct atgggcggcc gttaattaag cttttgttcc ctttagtgag ggttaattgc
     6481 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6541 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6601 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6661 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6721 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6781 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6841 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6901 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6961 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7021 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7081 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7141 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7201 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7261 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7321 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7381 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7441 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7501 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7561 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7621 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7681 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7741 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7801 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7861 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7921 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7981 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8041 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8101 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8161 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8221 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8281 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8341 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8401 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8461 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8521 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8581 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8641 agtgccac
//
