LOCUS       V026808                 8069 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026808
VERSION     V026808
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             896..3556
                     /label="Nedd4(NM_010890)"
                     /note="Nedd4(NM_010890)"
                     /gene="Nedd4"
     CDS             3572..3589
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    3636..3860
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3906..4334
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4348..4677
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4744..5538
                     /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    5712..5833
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5882..5898)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5906..5922
                     /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(5930..5960)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5975..5996
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6284..6869)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7040..7900)
                     /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(7901..8005)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg ccaccatgag
      901 ctcggacatg gcagccgacg agtcggaggc cccagtactc tcggaggacg aggtatggga
      961 gttttgcctg gataagacag aagatggtgg cggatccccc ggaagtgatg ttacagatac
     1021 ttgtgagcct ccatgtggat gctgggagtt gaatccgaat tccctggaag aggagcacgt
     1081 gctgttcact gctgatccgt acctggagct ccacaacgat gacacacgag ttgtgagagt
     1141 gaaggttata gctggcatag gcctggccaa gaaagacatc ttgggagcca gtgatcctta
     1201 cgtaagagtg acattgtatg acccgatgag tggaatcctt accagcgtgc agacaaaaac
     1261 tatcaaaaag tctttgaatc caaaatggaa tgaagaaata ctgttcaggg tccttccaca
     1321 gcgacaccgc attcttttcg aagtgtttga tgaaaatcgt ttgacaagag atgatttcct
     1381 aggtcaagtg gatgtccctc tctatccttt accgactgaa aacccaagaa tggagagacc
     1441 atatacattt aaggattttg ttcttcatcc aagaagtcac aaatcaagag ttaaaggtta
     1501 tctgagatta aaaatgactt atttacctaa aaatggctca gaagatgaaa atgcagacca
     1561 ggctgaggag ttagagcctg gctgggttgt tttggaccaa ccagatgctg ccactcattt
     1621 gccgcatcca ccagaaccct ctcccctacc tccaggatgg gaagagaggc aggatgtcct
     1681 tggaaggacc tactacgtaa accatgaatc tagaagaaca cagtggaaaa ggccaagccc
     1741 tgacgatgac ctcacggatg aagacaatga tgatatgcag ctgcaagcgc agcgagcatt
     1801 caccaccagg cggcagatat cggaggatgt ggatggccct gacaaccggg agtcccctga
     1861 gaattgggaa atcgtacgag aagatgaaaa caccgagtat agtggtcagg ctgtccagtc
     1921 acctccatcg ggtcacattg atgtgcagac tcaccttgca gaagagttta ataccagact
     1981 tgctgtgtgt ggaaatccag ccaccagcca gccggttacc agctcaaatc attccagcag
     2041 aggaggcagc ttgcagacct gtatctttga ggaacagcct acacttcctg tgcttttgcc
     2101 tacttcatct ggattgccac caggttggga agaaaaacaa gatgacagag gaagatcata
     2161 ctatgtagac cacaactcta aaaccaccac atggtccaag cccaccatgc aggatgatcc
     2221 aagatcgaaa atccctgctc atctgagagg aaagactgac tccaatgacc tgggaccctt
     2281 acctccaggc tgggaagaaa gaacccacac agatgggcga gtcttcttca taaaccacaa
     2341 tataaagaag acccagtggg aagatcctcg cctgcagaac gtggcaatca ctggaccagc
     2401 agtgccctac tccagagatt acaagagaaa gtacgagttc ttcagaagga agctcaagaa
     2461 gcagactgac attccaaaca aatttgaaat gaagcttcgc cgcgcaaaca ttctggagga
     2521 ttcttaccgg aggattatgg gtgtgaagag agctgacttg ctcaaggcca gactctggat
     2581 tgagtttgat ggtgaaaagg gccttgacta tggaggggtt gccagagagt ggttcttcct
     2641 catctcgaag gaaatgttca acccttacta cggcctgttt gaatattctg ctacggataa
     2701 ttacacccta cagataaatc ctaactcggg cttgtgtaat gaagatcacc tctcatactt
     2761 caagttcatt ggccgtgtgg ctgggatggc agtttatcat ggcaagctgt tggatgggtt
     2821 tttcatccgt ccgttttaca agatgatgct tcagaaactg ataacactgc acgacatgga
     2881 gtccgtggat agtgaatatt acagttctct gcgatggatt cttgaaaatg acccgacgga
     2941 gctggacctg agatttatca tagatgaaga actttttgga cagacacatc agcacgaact
     3001 gaaaaccgga ggatcagaga ttgttgtcac caataagaac aaaaaggagt atatctacct
     3061 tgtaatacaa tggcgatttg tgaaccgtat ccagaagcaa atggcagctt ttaaagaggg
     3121 attttttgaa ctgataccac aggatctcat caagatattt gatgaaaatg agctagagct
     3181 tctcatgtgt ggtctgggag atgtggatgt gaacgactgg cgggaacaca caaaatacaa
     3241 aaatggctac agcatgaacc accaggtcat ccactggttc tggaaggctg tttggatgat
     3301 ggattcggaa aaaagaatac gcttacttca gtttgtcact ggcacatccc gtgtgccgat
     3361 gaatgggttt gctgaactct atggctcgaa tggaccacaa tccttcacag tggaacaatg
     3421 gggcacccct gataagctgc caagagcaca cacctgcttc aatcgcctgg acctgccacc
     3481 ctacgaatcc tttgacgaac tctgggataa acttcagatg gcaattgaga acacacaggg
     3541 ctttgatggc gttgatggcg gaggtggcag tcatcatcac catcatcact aactcgagtc
     3601 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3661 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3721 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3781 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3841 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3901 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3961 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4021 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4081 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4141 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4201 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4261 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4321 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4381 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4441 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4501 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4561 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4621 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4681 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4741 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4801 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4861 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4921 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4981 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5041 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5101 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5161 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5221 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5281 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5341 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5401 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5461 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5521 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5581 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5641 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5701 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5761 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5821 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5881 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5941 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6001 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6061 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6121 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6181 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6241 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6301 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6361 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6421 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6481 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6541 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6601 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6661 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6721 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6781 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     6841 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6901 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6961 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7021 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7081 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7141 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7201 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7261 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7321 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7381 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7441 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7501 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7561 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7621 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7681 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7741 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7801 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7861 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7921 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7981 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     8041 gatcgggaga tctcccgatc ccctatggt
//
