LOCUS       V014250                 8335 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014250
VERSION     V014250
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8335)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..233
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             234..414
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    458..583
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1076..1309
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1493..1537
                     /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             1686..1727
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    1798..1915
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2005..2208
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2384..2449
                     /label="3xFLAG"
                     /note="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
     promoter        2494..2705
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             2718..2986
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3042..3707
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             3777..3830
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             3831..4427
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4431..5018
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5092..5325
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5397..5531
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5537..5672
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5710..5726)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5734..5750)
                     /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(5758..5788)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5803..5824)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6112..6700)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6874..7731)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7732..7836)
                     /label="AmpR promoter"
     primer_bind     8310..8326
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatgc tgtgctgtat gagaagaacc aaacaggttg aaaagaatga
     2341 tgaggaccaa aagatcctcg agcatcgaga tatctacgga tccgactaca aagaccatga
     2401 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt gagcggccgc
     2461 gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     2521 ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg
     2581 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     2641 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     2701 cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag
     2761 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa
     2821 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc
     2881 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc
     2941 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg
     3001 cgcctacgct agacgccacc atggagagcg acgagagcgg cctgcccgcc atggagatcg
     3061 agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc ggcggagagg
     3121 gcacccccaa gcagggccgc atgaccaaca agatgaagag caccaaaggc gccctgacct
     3181 tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc ggcacctacc
     3241 ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac accaacaccc
     3301 gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac cgctacgagg
     3361 ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag gacagcgtga
     3421 tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac cccatgggcg
     3481 ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc ggctactaca
     3541 gcttcgtggt ggacagccac atgcacttca agagcgccat ccaccccagc atcctgcaga
     3601 acgggggccc catgttcgcc ttccgccgcg tggaggagct gcacagcaac accgagctgg
     3661 gcatcgtgga gtaccagcac gccttcaaga cccccatcgc cttcgccaga tcccgcgctc
     3721 agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg ctccaccgga tctcgcgagg
     3781 gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct atgaccgagt
     3841 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     3901 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     3961 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4021 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4081 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4141 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4201 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4261 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     4321 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     4381 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga aatcaacctc
     4441 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     4501 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     4561 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     4621 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     4681 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     4741 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     4801 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     4861 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     4921 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc cgccttcgcc
     4981 ctcagacgag tcggatctcc ctttggccgc ctccccgcct ggtaccttta agaccaatga
     5041 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     5101 taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct ctctggttag
     5161 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     5221 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     5281 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     5341 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     5401 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5461 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5521 atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5581 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5641 ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg gcccaaattc
     5701 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5761 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5821 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5881 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5941 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6001 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6061 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6121 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6181 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6241 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6301 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6361 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6421 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6481 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6541 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6601 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6661 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6721 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6781 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6841 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6901 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6961 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7021 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7081 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7141 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7201 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7261 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7321 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7381 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7441 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7501 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7561 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7621 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7681 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7741 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7801 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7861 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     7921 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     7981 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     8041 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     8101 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     8161 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     8221 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     8281 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg
//
