LOCUS       V014295                 9555 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014295
VERSION     V014295
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9555)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..507
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             529..2178
                     /label="luciferase"
                     /note="firefly luciferase"
     CDS             2179..2244
                     /label="F2A"
                     /note="2A peptide from foot-and-mouth disease virus
                     polyprotein"
     CDS             2245..2841
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             2972..3205
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3277..3411
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      3438..3573
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(3594..3612)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3622..3638)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      3780..4235
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4261..4365
                     /label="AmpR promoter"
     CDS             4366..5223
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5397..5985
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    6273..6294
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6309..6339
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    6347..6363
                     /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     6371..6387
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        6408..6426
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6454..6680
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6681..6861
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6908..7033
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7526..7759
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             7944..7988
                     /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             8137..8178
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     promoter        8286..8526
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     CDS             complement(8595..8897)
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     promoter        complement(9249..9279)
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     misc_feature    9386..9503
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        9552..9555
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatccaccgg agcttaccat ggaagatgcc
      541 aaaaacatta agaagggccc agcgccattc tacccactcg aagacgggac cgccggcgag
      601 cagctgcaca aagccatgaa gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac
      661 gcacatatcg aggtggacat tacctacgcc gagtacttcg agatgagcgt tcggctggca
      721 gaagctatga agcgctatgg gctgaataca aaccatcgga tcgtggtgtg cagcgagaat
      781 agcttgcagt tcttcatgcc cgtgttgggt gccctgttca tcggtgtggc tgtggcccca
      841 gctaacgaca tctacaacga gcgcgagctg ctgaacagca tgggcatcag ccagcccacc
      901 gtcgtattcg tgagcaagaa agggctgcaa aagatcctca acgtgcaaaa gaagctaccg
      961 atcatacaaa agatcatcat catggatagc aagaccgact accagggctt ccaaagcatg
     1021 tacaccttcg tgacttccca tttgccaccc ggcttcaacg agtacgactt cgtgcccgag
     1081 agcttcgacc gggacaaaac catcgccctg atcatgaaca gtagtggcag taccggattg
     1141 cccaagggcg tagccctacc gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac
     1201 cccatcttcg gcaaccagat catccccgac accgctatcc tcagcgtggt gccatttcac
     1261 cacggcttcg gcatgttcac cacgctgggc tacttgatct gcggctttcg ggtcgtgctc
     1321 atgtaccgct tcgaggagga gctattcttg cgcagcttgc aagactataa gattcaatct
     1381 gccctgctgg tgcccacact atttagcttc ttcgctaaga gcactctcat cgacaagtac
     1441 gacctaagca acttgcacga gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt
     1501 gaggccgtgg ccaaacgctt ccacctacca ggcatccgcc agggctacgg cctgacagaa
     1561 acaaccagcg ccattctgat cacccccgaa ggggacgaca agcctggcgc agtaggcaag
     1621 gtggtgccct tcttcgaggc taaggtggtg gacttggaca caggtaagac actgggtgtg
     1681 aaccagcgcg gcgagctgtg cgtccgtggc cccatgatca tgagcggcta cgttaacaac
     1741 cccgaggcta caaacgctct catcgacaag gacggctggc tgcacagcgg cgacatcgcc
     1801 tactgggacg aggacgagca cttcttcatc gtggaccggc tgaagagcct gatcaaatac
     1861 aagggctacc aggtagcccc agccgaactg gagagcatcc tgctgcaaca ccccaacatc
     1921 ttcgacgccg gggtcgccgg cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc
     1981 gtcgtgctgg aacacggtaa aaccatgacc gagaaggaga tcgtggacta tgtggccagc
     2041 caggttacaa ccgccaagaa gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa
     2101 ggactgaccg gcaagttgga cgcccgcaag atccgcgaga ttctcattaa ggccaagaag
     2161 ggcggcaaga tcgccgtggt gaagcagacc ctgaactttg acctgctgaa gctggctggc
     2221 gatgtggagt ccaaccctgg ccccatgacc gagtacaagc ccacggtgcg cctcgccacc
     2281 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     2341 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     2401 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     2461 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     2521 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     2581 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     2641 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     2701 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     2761 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     2821 atgacccgca agcccggtgc ctgaggtacc cgcccgcccc acgacccgca gcgcccgacc
     2881 gaaaggagcg cacgacccca tgcatcttta agaccaatga cttacaaggc agctgtagat
     2941 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     3001 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     3061 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     3121 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     3181 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     3241 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     3301 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3361 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     3421 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3481 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3541 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     3601 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     3661 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     3721 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     3781 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3841 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3901 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3961 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4021 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4081 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4141 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4201 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     4261 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4321 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4381 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4441 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4501 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4561 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4621 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4681 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4741 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4801 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4861 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4921 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4981 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5041 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5101 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5161 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5221 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5281 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5341 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5401 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5461 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5521 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5581 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5641 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5701 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5761 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5821 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5881 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5941 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6001 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6061 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6121 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6181 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     6241 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     6301 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     6361 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     6421 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt
     6481 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
     6541 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
     6601 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
     6661 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct
     6721 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6781 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6841 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
     6901 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
     6961 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
     7021 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
     7081 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
     7141 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     7201 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     7261 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     7321 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     7381 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     7441 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     7501 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     7561 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     7621 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     7681 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     7741 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     7801 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     7861 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     7921 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     7981 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     8041 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     8101 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     8161 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
     8221 gattagtgaa cggatctcga cggtatcgat cacgagacta gcctcgagcg gccgccccct
     8281 tcaccgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     8341 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     8401 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     8461 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     8521 aaggacgaaa caccggtcgc gtgtcgccct ttgaagagca ttccgatagt ggctagcgac
     8581 tcgcctgaca tttatattcc ccagaacatc aggttaatgg cgtttttgat gtcattttcg
     8641 cggtggctga gatcagccac ttcttccccg ataacggaga ccggcacact ggccatatcg
     8701 gtggtcatca tgcgccagct ttcatccccg atatgcacca ccgggtaaag ttcacgggag
     8761 actttatctg acagcagacg tgcactggcc agggggatca ccatccgtcg cccgggcgtg
     8821 tcaataatat cactctgtac atccacaaac agacgataac ggctctctct tttataggtg
     8881 taaaccctaa actgcatttc accagcccct gttctcgtca gcaaaagagc cgttcatttc
     8941 aataaaccgg gcgacctcag ccatcccttc ctgattttcc gctttccagc gttcggcacg
     9001 cagacgacgg gcttcattct gcatggttgt gcttaccaga ccggagatat tgacatcata
     9061 tatgccttga gcaactgata gctgtcgctg tcaactgtca ctgtaatacg ctgcttcata
     9121 gcatacctct ttttgacata cttcgggtat acatatcagt atatattctt ataccgcaaa
     9181 aatcagcgcg caaatacgca tactgttatc tggcttttag taagccggat cctaactcaa
     9241 aatccacaca ttatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatggagt
     9301 cgcggccgcc actatcggaa tgctcttcaa agggcgacac gcgagaattc tcgacctcga
     9361 gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg
     9421 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     9481 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc
     9541 cgcggctcga ggggg
//
