LOCUS       V037520                 7350 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037520
VERSION     V037520
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             944..2020
                     /label="Hfe(NM_010424)"
                     /note="Hfe(NM_010424)"
                     /gene="Hfe"
     CDS             2033..2098
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2109..2731
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        2773..3070
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3124..3918
                     /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    4570..4704
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(4710..4728)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(4742..4758)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4766..4782
                     /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(4790..4820)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4835..4856
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5144..5732)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5903..6763)
                     /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(6764..6868)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6895..7323
                     /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 attaagcttg cggccgcgcc accatgagcc tatcagctgg
      961 gctccctgtg cggccgctgc tgctgctgct gctactgctg tggtccgtgg ccccgcaggc
     1021 actgccaccg cgttcacatt ctctaagata cctcttcatg ggtgcctcag agccagacct
     1081 cgggctgcct ttgtttgagg ctaggggcta tgtggatgac cagctctttg tgtcctacaa
     1141 tcatgagagt cgccgtgctg agcccagggc cccgtggatc ttggagcaaa cctcaagcca
     1201 gctgtggctg catctgagtc agagcctgaa agggtgggac tacatgttca tagtagactt
     1261 ctggaccatc atgggcaact ataaccacag taaggtcacg aagttgggag tggtgtccga
     1321 gtcccacatc ctgcaggtgg tcctaggctg tgaggtgcat gaagacaaca gtaccagcgg
     1381 cttctggaga tatggttatg acgggcaaga tcacctggaa ttctgcccca agacactaaa
     1441 ctggagcgca gccgagccag gggcctgggc caccaaggtg gaatgggacg agcacaagat
     1501 ccgtgccaaa cagaacaggg actacctgga gaaggactgc cccgagcagc tgaaacggct
     1561 cctggagctg gggagaggcg ttctgggaca gcaagtgcct actttggtga aagtgactcg
     1621 ccactgggcc tctacgggga cctctctaag gtgtcaggct ctggacttct tcccccagaa
     1681 catcactatg aggtggttga aggacaacca accactggat gccaaagatg tcaaccccga
     1741 gaaggtgcta cctaacgggg atgagaccta tcaaggctgg ctgacattgg ccgtggcccc
     1801 tggggacgag acaaggttca cctgtcaagt ggagcaccca ggcctggacc agcctctcac
     1861 tgcctcttgg gagcccttgc aatctcaggc catgattatc ggaatcatca gtggagtcac
     1921 cgtctgtgcc atcttcttgg ttggaattct gttcctaatc ttaaggaaaa ggaaggcttc
     1981 aggaggaacc atgggtggct atgtcttaac agactgtgag ggatcccggg ctgactacaa
     2041 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     2101 gtgatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     2161 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     2221 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     2281 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     2341 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     2401 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     2461 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     2521 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     2581 tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac
     2641 ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg
     2701 ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg
     2761 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2821 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2881 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2941 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3001 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3061 gcttttttgg aggaattgat cagcttggga tctgatcaag agacaggatg aggatcgttt
     3121 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3181 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3241 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3301 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3361 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3421 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3481 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3541 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3601 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3661 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3721 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3781 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3841 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3901 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     3961 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4021 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4081 tcgcccaccc cgggctcgat cccctcgcga gttggttcag ctgctgcctg aggctggacg
     4141 acctcgcgga gttctaccgg cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct
     4201 atccgcgcat ccatgccccc gaactgcagg agtggggagg cacgatggcc gctttggtcg
     4261 acccggacgg gacgctcctg cgcctgatac agaacgaatt gcttgcaggc atctcatgag
     4321 tgtgtcttcc cgttttccgc ctgaggtcac tgcgtggatg gagcgctggc gcctgctgcg
     4381 cgacggcgag ctgctcacca cccactcgcc aagctggaac cgtaaaaagg ccgcgttgct
     4441 ggcgtttttc cataggctcc gccgatcata atcagccata ccacatttgt agaggtttta
     4501 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     4561 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4621 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4681 aatgtatctt atcatgtctg gatcaattcc ctatagtgag tcgtattaaa ttcgtaatca
     4741 tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4801 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4861 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4921 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4981 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5041 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5101 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5161 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5221 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5281 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5341 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5401 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5461 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5521 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5581 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5641 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5701 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5761 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5821 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5881 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5941 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6001 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6061 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6121 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6181 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6241 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6301 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6361 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6421 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6481 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6541 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6601 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6661 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6721 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6781 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6841 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
     6901 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     6961 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     7021 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     7081 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     7141 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     7201 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     7261 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     7321 attttaacaa aatattaacg cttacaattt
//
