LOCUS       V033069                 7742 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033069
VERSION     V033069
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     CDS             8..31
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             47..2575
                     /label="FBXO11(NM_025133)"
                     /note="FBXO11(NM_025133)"
                     /gene="FBXO11"
     CDS             2627..3145
                     /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
                     TLKFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNG
                     IKANFKIRHNIE"
                     /label="VN173"
     polyA_signal    3156..3778
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        3807..4136
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(4175..4193)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(4207..4223)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4231..4247
                     /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(4255..4285)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4300..4321
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4609..5197)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5368..6228)
                     /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(6229..6333)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6360..6815
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     primer_bind     6956..6972
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        7133..7512
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        7513..7716
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
ORIGIN
        1 taccatggac tacaaagacg atgacgacaa gcttgcggcc gcgaatatgg ttgcagaaga
       61 atcaggtcct ggtgcacaaa atagtccata ccaacttcgt agaaaaactc ttttgccgaa
      121 aagaacagcg tgtcccacaa agaacagtat ggagggcgcc tcaacttcaa ctacagaaaa
      181 ctttggtcat cgtgcaaaac gtgcaagagt gtctggaaaa tcacaagatc tatcagcagc
      241 acctgctgaa cagtatcttc aggagaaact gccagatgaa gtggttctaa aaatcttctc
      301 ttacttgctg gaacaggatc tttgtagagc agcttgtgta tgtaaacgct tcagtgaact
      361 tgctaatgat ccaattttgt ggaaacgatt atatatggaa gtatttgaat atactcgccc
      421 tatgatgcat cctgaacctg gaaaattcta ccagattaat ccagaagagt atgaacatcc
      481 aaatccctgg aaagagagtt tccagcagtt gtataaaggt gcacatgtaa agccaggatt
      541 tgctgaacat ttctacagta accctgcaag atataaagga agagaaaata tgttgtatta
      601 tgatactatt gaagatgccc ttggtggggt acaagaggct cattttgatg gacttatctt
      661 tgttcattct ggaatatata ctgatgaatg gatatatatt gaatctccaa tcaccatgat
      721 tggtgcagca cctgggaaag tggcagacaa agttataatt gaaaacacta gagattcaac
      781 cttcgttttt atggaaggct ctgaagatgc ttatgttgga tatatgacaa taaggtttaa
      841 ccctgatgac aaatctgcac aacaccacaa tgcacaccac tgcttagaga ttacagtaaa
      901 ttgtagccct attattgatc actgtatcat ccgaagtaca tgtacagttg gttctgcagt
      961 atgtgttagt ggtcaaggag catgtcccac catcaagcac tgtaacatca gtgactgtga
     1021 aaatgttgga ctatatataa cagatcatgc acagggaata tatgaggata atgaaatttc
     1081 caataatgcg ttagctggga tttgggttaa aaatcatgga aacccaatta ttagacggaa
     1141 tcatattcat catggacgtg atgttggtgt gttcacattt gatcatggca tgggttactt
     1201 tgaaagttgc aatatacaca gaaataggat agcaggcttt gaagtaaaag cctatgctaa
     1261 ccctacagtg gttcgatgtg aaattcacca tgggcagact ggaggaatat atgtccatga
     1321 aaaaggaaga ggacaattca tagagaataa aatctatgca aacaactttg caggtgtatg
     1381 gattacctca aatagtgacc caacaataag gggaaattct atatttaatg gaaatcaagg
     1441 aggagtttac atctttggtg atggacgagg ccttattgaa ggaaatgaca tttatggcaa
     1501 tgcattagca ggaattcaaa ttaggacaaa cagttgtcca attgttcggc ataacaaaat
     1561 tcatgatggc cagcatggtg ggatttatgt gcatgaaaag ggacaaggag taatagaaga
     1621 gaatgaagtt tatagtaaca ctctagctgg agtctgggtg acaactggca gcactccagt
     1681 actgagaaga aaccggatac acagtggcaa gcaggttggt gtttattttt atgacaatgg
     1741 acatggagtg ctagaagaca atgatatcta taatcatatg tattcagggg ttcagataag
     1801 gactggaagc aaccccaaaa ttagacgcaa caaaatctgg ggaggacaga atggtggaat
     1861 tctagtttat aattctggtc taggctgtat agaagacaat gaaatatttg acaatgcaat
     1921 ggctggagtc tggattaaga cagatagtaa tcctacacta agaagaaata aaatccatga
     1981 tggaagagat ggtggcatct gtatatttaa tgggggtcga ggtctccttg aagaaaatga
     2041 tattttcagg aatgctcaag caggtgttct catcagcact aatagtcatc caatcttaag
     2101 gaaaaacaga atatttgatg gatttgccgc aggtattgaa attacaaatc acgcaactgc
     2161 aacactagaa ggcaatcaga tttttaacaa ccggtttgga ggcttatttt tagcatctgg
     2221 tgttaatgtg acaatgaaag ataacaaaat aatgaacaat caagatgcca tagaaaaggc
     2281 tgttagtaga ggccaatgtt tatataaaat atcaagttat accagctatc ccatgcatga
     2341 tttctacaga tgtcatactt gtaacaccac agatcgaaat gccatatgtg tgaactgcat
     2401 taagaagtgc catcagggac atgatgtaga gtttattaga catgataggt ttttctgtga
     2461 ctgtggtgct ggaacactgt ctaatccttg tacattagct ggtgagccta cacatgatac
     2521 agatacacta tatgactctg ctccacctat agaatctaat acattgcagc acaacggagg
     2581 tggaggttca ggtggtggag gatcttctag aagatccagc gccaccatgg tgagcaaggg
     2641 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2701 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2761 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2821 gacctggggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     2881 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2941 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3001 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3061 cgccatcagc gacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     3121 cttcaagatc cgccacaaca tcgagtaggg atcccgggtg gcatccctgt gacccctccc
     3181 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     3241 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     3301 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     3361 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     3421 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     3481 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     3541 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     3601 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg
     3661 aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga catttgagtt
     3721 gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg
     3781 gtacgcggcc agcttggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3841 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3901 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3961 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4021 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4081 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4141 tcctcgagga actgaaaaac cagaaagtta attccctata gtgagtcgta ttaaattcgt
     4201 aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4261 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4321 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4381 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4441 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4501 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4561 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4621 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4681 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4741 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4801 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4861 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4921 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4981 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5041 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5101 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5161 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5221 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5281 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5341 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5401 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5461 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5521 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5581 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5641 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5701 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5761 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5821 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5881 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5941 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6001 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6061 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6121 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6181 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6241 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6301 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6361 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     6421 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     6481 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     6541 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     6601 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     6661 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     6721 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     6781 cgcgaatttt aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac
     6841 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     6901 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     6961 acgacggcca gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta
     7021 gtcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta
     7081 tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga
     7141 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     7201 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     7261 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     7321 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     7381 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     7441 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc
     7501 gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac
     7561 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     7621 aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt
     7681 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaattga
     7741 tc
//
