LOCUS       V042684                 6410 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042684
VERSION     V042684
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             962..2200
                     /label="ADRB2(NM_000024)"
                     /note="ADRB2(NM_000024)"
                     /gene="ADRB2"
     CDS             2222..2740
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNG
                     IKANFKIRHNIE"
                     /label="VN173"
     polyA_signal    2751..3373
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        3402..3731
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(3770..3788)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(3802..3818)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3826..3842
                     /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(3850..3880)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    3895..3916
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4204..4792)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4963..5823)
                     /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(5824..5928)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5955..6383
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tatggggcaa cccgggaacg gcagcgcctt cttgctggca cccaatggaa gccatgcgcc
     1021 ggaccacgac gtcacgcagg aaagggacga ggtgtgggtg gtgggcatgg gcatcgtcat
     1081 gtctctcatc gtcctggcca tcgtgtttgg caatgtgctg gtcatcacag ccattgccaa
     1141 gttcgagcgt ctgcagacgg tcaccaacta cttcatcact tcactggcct gtgctgatct
     1201 ggtcatgggc ctggcagtgg tgccctttgg ggccgcccat attcttatga aaatgtggac
     1261 ttttggcaac ttctggtgcg agttttggac ttccattgat gtgctgtgcg tcacggccag
     1321 cattgagacc ctgtgcgtga tcgcagtgga tcgctacttt gccattactt cacctttcaa
     1381 gtaccagagc ctgctgacca agaataaggc ccgggtgatc attctgatgg tgtggattgt
     1441 gtcaggcctt acctccttct tgcccattca gatgcactgg taccgggcca cccaccagga
     1501 agccatcaac tgctatgcca atgagacctg ctgtgacttc ttcacgaacc aagcctatgc
     1561 cattgcctct tccatcgtgt ccttctacgt tcccctggtg atcatggtct tcgtctactc
     1621 cagggtcttt caggaggcca aaaggcagct ccagaagatt gacaaatctg agggccgctt
     1681 ccatgtccag aaccttagcc aggtggagca ggatgggcgg acggggcatg gactccgcag
     1741 atcttccaag ttctgcttga aggagcacaa agccctcaag acgttaggca tcatcatggg
     1801 cactttcacc ctctgctggc tgcccttctt catcgttaac attgtgcatg tgatccagga
     1861 taacctcatc cgtaaggaag tttacatcct cctaaattgg ataggctatg tcaattctgg
     1921 tttcaatccc cttatctact gccggagccc agatttcagg attgccttcc aggagcttct
     1981 gtgcctgcgc aggtcttctt tgaaggccta tgggaatggc tactccagca acggcaacac
     2041 aggggagcag agtggatatc acgtggaaca ggagaaagaa aataaactgc tgtgtgaaga
     2101 cctcccaggc acggaagact ttgtgggcca tcaaggtact gtgcctagcg ataacattga
     2161 ttcacaaggg aggaattgta gtacaaatga ctcactgctg tctagaagat ccatcgccac
     2221 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2281 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2341 cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac
     2401 cctcgtgacc accctgggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa
     2461 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2521 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2581 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2641 caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa
     2701 cggcatcaag gccaacttca agatccgcca caacatcgag tagggatccc gggtggcatc
     2761 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     2821 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     2881 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     2941 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     3001 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     3061 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     3121 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     3181 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat
     3241 aactatacca gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg
     3301 tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga
     3361 tgcctgttga attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg
     3421 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3481 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3541 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3601 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3661 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3721 gcttttgcaa aaagctcctc gaggaactga aaaaccagaa agttaattcc ctatagtgag
     3781 tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3841 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3901 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3961 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4021 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4081 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4141 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4201 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4261 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4321 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4381 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4441 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4501 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4561 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4621 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4681 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4741 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4801 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4861 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4921 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4981 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5041 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5101 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5161 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5221 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5281 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5341 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5401 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5461 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5521 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5581 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5641 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5701 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5761 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5821 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5881 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5941 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     6001 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     6061 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6121 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6181 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6241 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     6301 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     6361 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
//
