LOCUS       V016716                 9463 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V016716
VERSION     V016716
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9463)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..131
                     /label="AmpR promoter"
     CDS             132..989
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1163..1751
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2039..2060
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2075..2105
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2113..2129
                     /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     2137..2153
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        2174..2192
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2220..2447
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2448..2628
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2672..2797
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    3290..3523
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             3707..3751
                     /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             3900..3941
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    4012..4129
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        4152..4455
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4456..4659
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    4764..4788
                     /label="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4800..4841
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     CDS             4917..6566
                     /label="luciferase"
                     /note="firefly luciferase"
     protein_bind    complement(6576..6600)
                     /label="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    6631..7219
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     promoter        7275..7605
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        7653..7700
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             7719..8114
                     /label="BSD"
                     /note="blasticidin S deaminase"
     LTR             8204..8437
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8509..8643
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      8649..8784
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(8817..8835)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8845..8861)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      9006..9461
                     /direction=RIGHT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
       61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
      541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
      601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
      661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
      721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
      781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
      841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
      901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
      961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     2161 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
     2221 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     2281 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
     2341 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
     2401 gcattgcaga gatattgtat ttaagtgcct agctcgatac aataaacggg tctctctggt
     2461 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     2521 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     2581 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     2641 cagggacctg aaagcgaaag ggaaaccaga gctctctcga cgcaggactc ggcttgctga
     2701 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     2761 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     2821 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     2881 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     2941 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     3001 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     3061 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     3121 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     3181 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     3241 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     3301 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcctcaatg acgctgacgg
     3361 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     3421 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     3481 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     3541 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     3601 tggaacagat tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     3661 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     3721 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     3781 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     3841 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     3901 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     3961 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggttaa cttttaaaag
     4021 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     4081 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta tcgataagct
     4141 tgggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     4201 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     4261 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     4321 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     4381 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     4441 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     4501 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     4561 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     4621 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc
     4681 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccgact ctagtccagt
     4741 gtggtggaat tctgcagata tcaacaagtt tgtacaaaaa agcaggctcc accatggaag
     4801 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gggatccatg ggaaccaatt
     4861 cagtccctag gcttttgcaa aaagcttggc attccggtac tgttggtaaa gccaccatgg
     4921 aagacgccaa aaacataaag aaaggcccgg cgccattcta tccgctggaa gatggaaccg
     4981 ctggagagca actgcataag gctatgaaga gatacgccct ggttcctgga acaattgctt
     5041 ttacagatgc acatatcgag gtggacatca cttacgctga gtacttcgaa atgtccgttc
     5101 ggttggcaga agctatgaaa cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca
     5161 gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc gttatttatc ggagttgcag
     5221 ttgcgcccgc gaacgacatt tataatgaac gtgaattgct caacagtatg ggcatttcgc
     5281 agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa
     5341 agctcccaat catccaaaaa attattatca tggattctaa aacggattac cagggatttc
     5401 agtcgatgta cacgttcgtc acatctcatc tacctcccgg ttttaatgaa tacgattttg
     5461 tgccagagtc cttcgatagg gacaagacaa ttgcactgat catgaactcc tctggatcta
     5521 ctggtctgcc taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg
     5581 ccagagatcc tatttttggc aatcaaatca ttccggatac tgcgatttta agtgttgttc
     5641 cattccatca cggttttgga atgtttacta cactcggata tttgatatgt ggatttcgag
     5701 tcgtcttaat gtatagattt gaagaagagc tgtttctgag gagccttcag gattacaaga
     5761 ttcaaagtgc gctgctggtg ccaaccctat tctccttctt cgccaaaagc actctgattg
     5821 acaaatacga tttatctaat ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg
     5881 aagtcgggga agcggttgcc aagaggttcc atctgccagg tatcaggcaa ggatatgggc
     5941 tcactgagac tacatcagct attctgatta cacccgaggg ggatgataaa ccgggcgcgg
     6001 tcggtaaagt tgttccattt tttgaagcga aggttgtgga tctggatacc gggaaaacgc
     6061 tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg tccggttatg
     6121 taaacaatcc ggaagcgacc aacgccttga ttgacaagga tggatggcta cattctggag
     6181 acatagctta ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg aagtctctga
     6241 ttaagtacaa aggctatcag gtggctcccg ctgaattgga atccatcttg ctccaacacc
     6301 ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg
     6361 ccgccgttgt tgttttggag cacggaaaga cgatgacgga aaaagagatc gtggattacg
     6421 tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag
     6481 taccgaaagg tcttaccgga aaactcgacg caagaaaaat cagagagatc ctcataaagg
     6541 ccaagaaggg cggaaagatc gccgtgtaat tctagaccca gctttcttgt acaaagtggt
     6601 tgatatccag cacagtggcg gccgctcgac aatcaacctc tggattacaa aatttgtgaa
     6661 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     6721 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     6781 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     6841 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     6901 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     6961 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     7021 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     7081 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     7141 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     7201 ctttgggccg cctccccgcc tggaattctg cagatatccg gttagtaatg agtttggaat
     7261 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc
     7321 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     7381 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     7441 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     7501 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     7561 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     7621 tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata
     7681 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagcct ttgtctcaag
     7741 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag
     7801 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
     7861 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
     7921 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
     7981 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag
     8041 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
     8101 atgtgtggga gggctaagca caattcgagc tcggtacctt taagaccaat gacttacaag
     8161 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctagctcac
     8221 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     8281 tgagcctggg agctctctgg ctgcctaggg aacccactgc ttaagcctca ataaagcttg
     8341 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     8401 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     8461 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     8521 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     8581 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     8641 ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     8701 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     8761 agtgaggagg cttttttgga ggcctaggct tttgcgtcga gacgtaccca attcgcccta
     8821 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     8881 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     8941 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     9001 gcgcgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     9061 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     9121 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     9181 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     9241 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     9301 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     9361 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     9421 atttaacgcg aattttaaca aaatattaac gtttacaatt tcc
//
