LOCUS       V031964                 8979 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031964
VERSION     V031964
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..204
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             293..1609
                     /label="TFAP2A(NM_001372066)"
                     /note="TFAP2A(NM_001372066)"
                     /gene="TFAP2A"
     CDS             1616..1681
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        1725..1936
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             1949..2217
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             2245..3039
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLAATGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    3046..3634
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             3708..3941
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    4013..4134
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      4174..4309
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(4342..4358)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4366..4382
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(4390..4420)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4435..4456
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4744..5332)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5503..6363)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(6364..6468)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     6942..6958
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        6973..7199
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             7200..7380
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    7427..7552
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    8045..8278
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             8463..8507
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    8774..8890
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggttt atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagattc tagagctagc gaattcgcca ccatgaaaat
      301 gctttggaaa ttgacggata atatcaagta cgaggactgc gaggaccgtc acgacggcac
      361 cagcaacggg acggcacggt tgccccagct gggcactgta ggtcaatctc cctacacgag
      421 cgccccgccg ctgtcccaca cccccaatgc cgacttccag cccccatact tccccccacc
      481 ctaccagcct atctaccccc agtcgcaaga tccttactcc cacgtcaacg acccctacag
      541 cctgaacccc ctgcacgccc agccgcagcc gcagcaccca ggctggcccg gccagaggca
      601 gagccaggag tctgggctcc tgcacacgca ccgggggctg cctcaccagc tgtcgggcct
      661 ggatcctcgc agggactaca ggcggcacga ggacctcctg cacggcccac acgcgctcag
      721 ctcaggactc ggagacctct cgatccactc cttacctcac gccatcgagg aggtcccgca
      781 tgtagaagac ccgggtatta acatcccaga tcaaactgta attaagaaag gccccgtgtc
      841 cctgtccaag tccaacagca atgccgtctc cgccatccct attaacaagg acaacctctt
      901 cggcggcgtg gtgaacccca acgaagtctt ctgttcagtt ccgggtcgcc tctcgctcct
      961 cagctccacc tcgaagtaca aggtcacggt ggcggaagtg cagcggcggc tctcaccacc
     1021 cgagtgtctc aacgcgtcgc tgctgggcgg agtgctccgg agggcgaagt ctaaaaatgg
     1081 aggaagatct ttaagagaaa aactggacaa aataggatta aatctgcctg cagggagacg
     1141 taaagctgcc aacgttaccc tgctcacatc actagtagag ggagaagctg tccacctagc
     1201 cagggacttt gggtacgtgt gcgaaaccga atttcctgcc aaagcagtag ctgaatttct
     1261 caaccgacaa cattccgatc ccaatgagca agtgacaaga aaaaacatgc tcctggctac
     1321 aaaacagata tgcaaagagt tcaccgacct gctggctcag gaccgatctc ccctggggaa
     1381 ctcacggccc aaccccatcc tggagcccgg catccagagc tgcttgaccc acttcaacct
     1441 catctcccac ggcttcggca gccccgcggt gtgtgccgcg gtcacggccc tgcagaacta
     1501 tctcaccgag gccctcaagg ccatggacaa aatgtacctc agcaacaacc ccaacagcca
     1561 cacggacaac aacgccaaaa gcagtgacaa agaggagaag cacagaaagc tcgaggacta
     1621 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     1681 gtaggcggcc gcaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc
     1741 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag
     1801 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     1861 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     1921 tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg
     1981 ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg
     2041 tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct
     2101 ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac
     2161 cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa
     2221 gctgtgaccg gcgcctacgt cgagatgatt gaacaagatg gattgcacgc aggttctccg
     2281 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2341 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2401 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccgcg
     2461 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     2521 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     2581 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     2641 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     2701 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     2761 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     2821 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     2881 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     2941 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3001 cgcatcgcct tctatcgcct tcttgacgag ttcttctgac tcgacaatca acctctggat
     3061 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3121 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3181 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3241 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3301 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3361 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3421 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     3481 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3541 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3601 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta
     3661 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     3721 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca
     3781 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     3841 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     3901 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     3961 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     4021 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4081 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4141 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4201 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4261 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc
     4321 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4381 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4441 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4501 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4561 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4621 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4681 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4741 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4801 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4861 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4921 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4981 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5041 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5101 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5161 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5221 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5281 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5341 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5401 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5461 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5521 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5581 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5641 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5701 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5761 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5821 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5881 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5941 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6001 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6061 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6121 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6181 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6241 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6301 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6361 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6421 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6481 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     6541 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     6601 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     6661 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     6721 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     6781 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     6841 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     6901 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     6961 caagctgacg cgtgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga
     7021 gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa
     7081 ggtggtacga tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa
     7141 ccactgaatt gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg
     7201 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     7261 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     7321 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     7381 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     7441 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     7501 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     7561 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     7621 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     7681 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     7741 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     7801 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     7861 aaacaaaagt aagaccaccg cacagcaagc ggccactgat cttcagacct ggaggaggag
     7921 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     7981 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     8041 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     8101 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     8161 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     8221 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     8281 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     8341 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     8401 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     8461 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     8521 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     8581 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     8641 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     8701 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
     8761 ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     8821 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat
     8881 tcaaaatttt atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact
     8941 tggcagtaca tctacgtatt agtcatcgct attaccatg
//
