LOCUS       V040888                 7590 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040888
VERSION     V040888
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             931..996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1000..2295
                     /label="AHCY(NM_000687)"
                     /note="AHCY(NM_000687)"
                     /gene="AHCY"
     polyA_signal    2349..2971
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        3013..3310
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3364..4158
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4810..4944
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(4950..4968)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(4982..4998)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5006..5022
                     /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(5030..5060)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5075..5096
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5384..5972)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6143..7003)
                     /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(7004..7108)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7135..7563
                     /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 attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tgtctgacaa actgccctac
     1021 aaagtcgccg acatcggcct ggctgcctgg ggacgcaagg ccctggacat tgctgagaac
     1081 gagatgccgg gcctgatgcg tatgcgggag cggtactcgg cctccaagcc actgaagggc
     1141 gcccgcatcg ctggctgcct gcacatgacc gtggagacgg ccgtcctcat tgagaccctc
     1201 gtcaccctgg gtgctgaggt gcagtggtcc agctgcaaca tcttctccac ccaggaccat
     1261 gcggcggctg ccattgccaa ggctggcatt ccggtgtatg cctggaaggg cgaaacggac
     1321 gaggagtacc tgtggtgcat tgagcagacc ctgtacttca aggacgggcc cctcaacatg
     1381 attctggacg acgggggcga cctcaccaac ctcatccaca ccaagtaccc gcagcttctg
     1441 ccaggcatcc gaggcatctc tgaggagacc acgactgggg tccacaacct ctacaagatg
     1501 atggccaatg ggatcctcaa ggtgcctgcc atcaatgtca atgactccgt caccaagagc
     1561 aagtttgaca acctctatgg ctgccgggag tccctcatag atggcatcaa gcgggccaca
     1621 gatgtgatga ttgccggcaa ggtagcggtg gtagcaggct atggtgatgt gggcaagggc
     1681 tgtgcccagg ccctgcgggg tttcggagcc cgcgtcatca tcaccgagat tgaccccatc
     1741 aacgcactgc aggctgccat ggagggctat gaggtgacca ccatggatga ggcctgtcag
     1801 gagggcaaca tctttgtcac caccacaggc tgtattgaca tcatccttgg ccggcacttt
     1861 gagcagatga aggatgatgc cattgtgtgt aacattggac actttgacgt ggagatcgat
     1921 gtcaagtggc tcaacgagaa cgccgtggag aaggtgaaca tcaagccgca ggtggaccgg
     1981 tatcggttga agaatgggcg ccgcatcatc ctgctggccg agggtcggct ggtcaacctg
     2041 ggttgtgcca tgggccaccc cagcttcgtg atgagtaact ccttcaccaa ccaggtgatg
     2101 gcgcagatcg agctgtggac ccatccagac aagtaccccg ttggggttca tttcctgccc
     2161 aagaagctgg atgaggcagt ggctgaagcc cacctgggca agctgaatgt gaagttgacc
     2221 aagctaactg agaagcaagc ccagtacctg ggcatgtcct gtgatggccc cttcaagccg
     2281 gatcactacc gctactgaga attcatcgat agatctgata tcggtaccag tcgactctag
     2341 aggatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     2401 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     2461 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     2521 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     2581 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     2641 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     2701 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     2761 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     2821 tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac
     2881 ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg
     2941 ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg
     3001 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3061 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3121 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3181 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3241 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3301 gcttttttgg aggaattgat cagcttggga tctgatcaag agacaggatg aggatcgttt
     3361 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3421 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3481 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3541 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3601 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3661 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3721 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3781 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3841 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3901 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3961 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4021 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4081 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4141 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4201 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4261 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4321 tcgcccaccc cgggctcgat cccctcgcga gttggttcag ctgctgcctg aggctggacg
     4381 acctcgcgga gttctaccgg cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct
     4441 atccgcgcat ccatgccccc gaactgcagg agtggggagg cacgatggcc gctttggtcg
     4501 acccggacgg gacgctcctg cgcctgatac agaacgaatt gcttgcaggc atctcatgag
     4561 tgtgtcttcc cgttttccgc ctgaggtcac tgcgtggatg gagcgctggc gcctgctgcg
     4621 cgacggcgag ctgctcacca cccactcgcc aagctggaac cgtaaaaagg ccgcgttgct
     4681 ggcgtttttc cataggctcc gccgatcata atcagccata ccacatttgt agaggtttta
     4741 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     4801 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4861 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4921 aatgtatctt atcatgtctg gatcaattcc ctatagtgag tcgtattaaa ttcgtaatca
     4981 tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5041 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5101 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5161 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5221 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5281 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5341 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5401 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5461 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5521 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5581 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5641 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5701 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5761 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5821 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5881 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5941 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6001 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6061 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6121 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6181 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6241 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6301 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6361 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6421 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6481 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6541 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6601 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6661 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6721 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6781 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6841 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6901 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6961 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7021 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7081 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
     7141 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     7201 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     7261 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     7321 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     7381 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     7441 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     7501 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     7561 attttaacaa aatattaacg cttacaattt
//
