LOCUS       V039976                 9817 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039976
VERSION     V039976
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"
     regulatory      2284..2293
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2293..2364
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags"
                     /transl_table=1
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
                     /label="3xFLAG"
     CDS             2371..4146
                     /label="CANX(NM_001746)"
                     /note="CANX(NM_001746)"
                     /gene="CANX"
     CDS             4201..4863
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     promoter        4982..5193
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5206..5474
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5507..5905
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5912..6500
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6574..6807
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6879..7000
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7019..7154
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7192..7208)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7216..7232
                     /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(7240..7270)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7285..7306
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7594..8182)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8353..9213)
                     /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(9214..9318)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9792..9808
                     /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 agagccacca tggactacaa ggatgacgat gacaaggatt acaaagacga cgatgataag
     2341 gactataagg atgatgacga caaagaattc atggaaggga agtggttgct gtgtatgtta
     2401 ctggtgcttg gaactgctat tgttgaggct catgatggac atgatgatga tgtgattgat
     2461 attgaggatg accttgacga tgtcattgaa gaggtagaag actcaaaacc agataccact
     2521 gctcctcctt catctcccaa ggttacttac aaagctccag ttccaacagg ggaagtatat
     2581 tttgctgatt cttttgacag aggaactctg tcagggtgga ttttatccaa agccaagaaa
     2641 gacgataccg atgatgaaat tgccaaatat gatggaaagt gggaggtaga ggaaatgaag
     2701 gagtcaaagc ttccaggtga taaaggactt gtgttgatgt ctcgggccaa gcatcatgcc
     2761 atctctgcta aactgaacaa gcccttcctg tttgacacca agcctctcat tgttcagtat
     2821 gaggttaatt tccaaaatgg aatagaatgt ggtggtgcct atgtgaaact gctttctaaa
     2881 acaccagaac tcaacctgga tcagttccat gacaagaccc cttatacgat tatgtttggt
     2941 ccagataaat gtggagagga ctataaactg cacttcatct tccgacacaa aaaccccaaa
     3001 acgggtatct atgaagaaaa acatgctaag aggccagatg cagatctgaa gacctatttt
     3061 actgataaga aaacacatct ttacacacta atcttgaatc cagataatag ttttgaaata
     3121 ctggttgacc aatctgtggt gaatagtgga aatctgctca atgacatgac tcctcctgta
     3181 aatccttcac gtgaaattga ggacccagaa gaccggaagc ccgaggattg ggatgaaaga
     3241 ccaaaaatcc cagatccaga agctgtcaag ccagatgact gggatgaaga tgcccctgct
     3301 aagattccag atgaagaggc cacaaaaccc gaaggctggt tagatgatga gcctgagtac
     3361 gtacctgatc cagacgcaga gaaacctgag gattgggatg aagacatgga tggagaatgg
     3421 gaggctcctc agattgccaa ccctagatgt gagtcagctc ctggatgtgg tgtctggcag
     3481 cgacctgtga ttgacaaccc caattataaa ggcaaatgga agcctcctat gattgacaat
     3541 cccagttacc agggaatctg gaaacccagg aaaataccaa atccagattt ctttgaagat
     3601 ctggaacctt tcagaatgac tccttttagt gctattggtt tggagctgtg gtccatgacc
     3661 tctgacattt tttttgacaa ctttatcatt tgtgctgatc gaagaatagt tgatgattgg
     3721 gccaatgatg gatggggcct gaagaaagct gctgatgggg ctgctgagcc aggcgttgtg
     3781 gggcagatga tcgaggcagc tgaagagcgc ccgtggctgt gggtagtcta tattctaact
     3841 gtagcccttc ctgtgttcct ggttatcctc ttctgctgtt ctggaaagaa acagaccagt
     3901 ggtatggagt ataagaaaac tgatgcacct caaccggatg tgaaggaaga ggaagaagag
     3961 aaggaagagg aaaaggacaa gggagatgag gaggaggaag gagaagagaa acttgaagag
     4021 aaacagaaaa gtgatgctga agaagatggt ggcactgtca gtcaagagga ggaagacaga
     4081 aaacctaaag cagaggagga tgaaattttg aacagatcac caagaaacag aaagccacga
     4141 agagagggag gagggggatc aggaggcggt ggatctatgg agagcgacga gagcggcctg
     4201 cccgccatgg agatcgagtg ccgcatcacc ggcaccctga acggcgtgga gttcgagctg
     4261 gtgggcggcg gagagggcac ccccaagcag ggccgcatga ccaacaagat gaagagcacc
     4321 aaaggcgccc tgaccttcag cccctacctg ctgagccacg tgatgggcta cggcttctac
     4381 cacttcggca cctaccccag cggctacgag aaccccttcc tgcacgccat caacaacggc
     4441 ggctacacca acacccgcat cgagaagtac gaggacggcg gcgtgctgca cgtgagcttc
     4501 agctaccgct acgaggccgg ccgcgtgatc ggcgacttca aggtggtggg caccggcttc
     4561 cccgaggaca gcgtgatctt caccgacaag atcatccgca gcaacgccac cgtggagcac
     4621 ctgcacccca tgggcgataa cgtgctggtg ggcagcttcg cccgcacctt cagcctgcgc
     4681 gacggcggct actacagctt cgtggtggac agccacatgc acttcaagag cgccatccac
     4741 cccagcatcc tgcagaacgg gggccccatg ttcgccttcc gccgcgtgga ggagctgcac
     4801 agcaacaccg agctgggcat cgtggagtac cagcacgcct tcaagacccc catcgccttc
     4861 gccagatccc gcgctcagtc gtccaattct gccgtggacg gcaccgccgg acccggctcc
     4921 accggatctc gctaggatcc gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     4981 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     5041 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     5101 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     5161 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     5221 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     5281 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     5341 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     5401 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     5461 cgccgttaca gatccaagct gtgaccggcg cctacgctag gccaccatgg ccaagccttt
     5521 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
     5581 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
     5641 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
     5701 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
     5761 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     5821 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     5881 ctctggttat gtgtgggagg gctaagtcga caatcaacct ctggattaca aaatttgtga
     5941 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     6001 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     6061 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     6121 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     6181 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     6241 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     6301 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     6361 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     6421 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     6481 cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag
     6541 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     6601 aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6661 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6721 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6781 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6841 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     6901 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6961 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     7021 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     7081 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     7141 cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct
     7201 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7261 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7321 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7381 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7441 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7501 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7561 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7621 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7681 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7741 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7801 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7861 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7921 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7981 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     8041 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8101 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     8161 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     8221 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8281 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8341 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8401 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8461 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8521 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8581 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8641 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8701 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8761 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8821 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8881 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8941 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     9001 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     9061 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     9121 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     9181 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     9241 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9301 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     9361 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     9421 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     9481 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     9541 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     9601 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     9661 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     9721 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     9781 gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
