LOCUS       V034179                 9976 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034179
VERSION     V034179
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..4830
                     /label="AXIN2(NM_004655)"
                     /note="AXIN2(NM_004655)"
                     /gene="AXIN2"
     CDS             4837..4902
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4946..5157
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5170..5438
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5465..6064
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    6071..6659
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6733..6966
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7038..7159
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7178..7313
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7351..7367)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7375..7391
                     /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(7399..7429)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7444..7465
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7753..8341)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8512..9372)
                     /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(9373..9477)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9951..9967
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
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 ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgagtagc gctatgttgg tgacttgcct cccggacccc
     2341 agcagcagct tccgtgagga tgccccgcgg cccccagtgc caggggaaga aggggagacc
     2401 ccaccgtgtc agccaggggt gggcaagggc caggtcacca aacccatgcc tgtctcttcc
     2461 aacaccaggc ggaacgaaga tgggttgggg gagccggagg ggcgggcatc tccggattcc
     2521 cctctgaccc ggtggaccaa gtccttacac tccttattgg gcgatcaaga cggtgcttac
     2581 ctgttccgaa ctttcctgga gagggagaaa tgcgtggata ccttagactt ctggtttgcc
     2641 tgcaatggat tcaggcagat gaacctgaag gataccaaaa ctttacgagt agccaaagcg
     2701 atctacaaaa ggtacattga gaacaacagc attgtctcca agcagctgaa gcctgccacc
     2761 aagacctaca taagagatgg catcaagaag cagcagattg attccatcat gtttgaccag
     2821 gcgcagaccg agatccagtc ggtgatggag gaaaatgcct accagatgtt tttgacttct
     2881 gatatatacc tcgaatatgt gaggagtggg ggagaaaaca cagcttacat gagtaatggg
     2941 ggactcggga gcctaaaggt cgtgtgtggc tatctcccca ccttgaatga agaagaggag
     3001 tggacttgtg ccgacttcaa gtgcaaactt tcgccaaccg tggttggctt gtccagcaaa
     3061 actctgaggg ccacggcgag tgtgaggtcc acggaaactg ttgacagtgg atacaggtcc
     3121 ttcaagagga gcgatcctgt taatccttat cacataggtt ctggctatgt ctttgcacca
     3181 gccaccagcg ccaacgacag tgagatatcc agtgatgcgc tgacggatga ttccatgtcc
     3241 atgacggaca gcagtgtaga tggaattcct ccttatcgtg tgggcagtaa gaaacagctc
     3301 cagagagaaa tgcatcgcag tgtgaaggcc aatggccaag tgtctctacc tcatttcccg
     3361 agaacccacc gcctgcccaa ggagatgacc cccgtggaac ccgccacctt tgcagctgag
     3421 ctgatctcga ggctggaaaa gctgaagctg gagttggaga gccgccacag cctggaggag
     3481 cgcctgcagc agatccgaga ggatgaagag agagagggct ccgagctcac actcaattcg
     3541 cgggaggggg cgcccacgca gcaccccctc tccctactgc cctccggcag ctacgaggaa
     3601 gacccgcaga cgatactgga cgatcacctg tccagggtcc tcaagacccc tggctgccag
     3661 tctccaggcg taggccgcta tagcccccgc tcccgctccc cggaccacca ccaccaccac
     3721 cattcgcagt accactccct gctcccgccc ggtggcaagc tgcctcccgc ggccgcctcg
     3781 ccgggcgcct gccccctcct cgggggcaaa ggctttgtga ccaagcagac gacgaagcat
     3841 gtccaccacc actacatcca ccaccatgcc gtccccaaga ccaaggagga gatcgaggcg
     3901 gaggccacgc agcgggtgca ctgcttctgc cctgggggca gcgagtatta ctgctactcg
     3961 aaatgcaaaa gccactccaa ggctccggaa accatgccca gcgagcagtt tggcggcagc
     4021 agaggcagta ccttgcccaa acgcaatggg aaaggcacgg agccgggcct ggccctgccc
     4081 gccagggaag gaggggcccc cggcggagct ggggccctgc agcttccccg ggaggaagga
     4141 gacaggtcgc aggatgtctg gcagtggatg ctggagagtg agcggcagag caagcccaag
     4201 ccccatagtg cccaaagcac aaaaaaggcc taccccttgg agtctgcccg ctcgtctcca
     4261 ggcgaacgag ccagccggca ccatctgtgg gggggcaaca gcgggcaccc ccgcaccacc
     4321 ccccgtgccc acctgttcac ccaggaccct gcgatgcctc ccctgacccc acccaacacg
     4381 ctggctcagc tggaggaggc ctgtcgcagg ctagctgagg tgtcgaagcc cccaaagcag
     4441 cggtgctgtg tggccagtca gcagagggac aggaatcatt cggccactgt tcagacggga
     4501 gccacaccct tctccaatcc aagcctggct ccagaagatc acaaagagcc aaagaaactg
     4561 gcaggtgtcc acgcgctcca ggccagtgag ttggttgtca cttacttttt ctgtggggaa
     4621 gaaattccat accggaggat gctgaaggct cagagcttga ccctgggcca ctttaaagag
     4681 cagctcagca aaaagggaaa ttataggtat tacttcaaaa aagcaagcga tgagtttgcc
     4741 tgtggagcgg tgtttgagga gatctgggag gatgagacgg tgctcccgat gtatgaaggc
     4801 cggattctgg gcaaagtgga gcggatcgat ggatccgact acaaagacca tgacggtgat
     4861 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc
     4921 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     4981 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg
     5041 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     5101 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg
     5161 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat
     5221 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc
     5281 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg
     5341 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac
     5401 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg
     5461 ctagatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc
     5521 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc
     5581 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     5641 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     5701 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     5761 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     5821 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     5881 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     5941 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     6001 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     6061 ctgagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     6121 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     6181 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     6241 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     6301 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     6361 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     6421 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     6481 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     6541 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     6601 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     6661 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     6721 aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt
     6781 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     6841 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     6901 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6961 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     7021 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     7081 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     7141 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag
     7201 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     7261 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt
     7321 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     7381 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7441 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7501 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7561 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7621 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7681 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7921 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     8041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     8101 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     8161 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     8221 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     8281 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     8341 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     8401 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     8461 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     8521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     8581 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8641 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8701 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8761 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8821 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8881 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8941 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     9001 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     9061 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     9121 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     9181 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     9241 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     9301 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     9361 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     9421 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     9481 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     9541 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     9601 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     9661 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     9721 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     9781 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     9841 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     9901 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     9961 ggccagtgcc aagctg
//
