LOCUS       V019989                 7589 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019989
VERSION     V019989
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             908..3025
                     /label="FZD6(NM_003506)"
                     /note="FZD6(NM_003506)"
                     /gene="FZD6"
     CDS             3032..3121
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    3156..3380
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3426..3854
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3868..4197
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4264..5058
                     /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    5232..5353
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5402..5418)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5426..5442
                     /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(5450..5480)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5495..5516
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5804..6389)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6560..7420)
                     /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(7421..7525)
                     /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 agctcggatc
      901 cgccaccatg gaaatgttta catttttgtt gacgtgtatt tttctacccc tcctaagagg
      961 gcacagtctc ttcacctgtg aaccaattac tgttcccaga tgtatgaaaa tggcctacaa
     1021 catgacgttt ttccctaatc tgatgggtca ttatgaccag agtattgccg cggtggaaat
     1081 ggagcatttt cttcctctcg caaatctgga atgttcacca aacattgaaa ctttcctctg
     1141 caaagcattt gtaccaacct gcatagaaca aattcatgtg gttccacctt gtcgtaaact
     1201 ttgtgagaaa gtatattctg attgcaaaaa attaattgac acttttggga tccgatggcc
     1261 tgaggagctt gaatgtgaca gattacaata ctgtgatgag actgttcctg taacttttga
     1321 tccacacaca gaatttcttg gtcctcagaa gaaaacagaa caagtccaaa gagacattgg
     1381 attttggtgt ccaaggcatc ttaagacttc tgggggacaa ggatataagt ttctgggaat
     1441 tgaccagtgt gcgcctccat gccccaacat gtattttaaa agtgatgagc tagagtttgc
     1501 aaaaagtttt attggaacag tttcaatatt ttgtctttgt gcaactctgt tcacattcct
     1561 tactttttta attgatgtta gaagattcag atacccagag agaccaatta tatattactc
     1621 tgtctgttac agcattgtat ctcttatgta cttcattgga tttttgctag gcgatagcac
     1681 agcctgcaat aaggcagatg agaagctaga acttggtgac actgttgtcc taggctctca
     1741 aaataaggct tgcaccgttt tgttcatgct tttgtatttt ttcacaatgg ctggcactgt
     1801 gtggtgggtg attcttacca ttacttggtt cttagctgca ggaagaaaat ggagttgtga
     1861 agccatcgag caaaaagcag tgtggtttca tgctgttgca tggggaacac caggtttcct
     1921 gactgttatg cttcttgcta tgaacaaagt tgaaggagac aacattagtg gagtttgctt
     1981 tgttggcctt tatgacctgg atgcttctcg ctactttgta ctcttgccac tgtgcctttg
     2041 tgtgtttgtt gggctctctc ttcttttagc tggcattatt tccttaaatc atgttcgaca
     2101 agtcatacaa catgatggcc ggaaccaaga aaaactaaag aaatttatga ttcgaattgg
     2161 agtcttcagc ggcttgtatc ttgtgccatt agtgacactt ctcggatgtt acgtctatga
     2221 gcaagtgaac aggattacct gggagataac ttgggtctct gatcattgtc gtcagtacca
     2281 tatcccatgt ccttatcagg caaaagcaaa agctcgacca gaattggctt tatttatgat
     2341 aaaatacctg atgacattaa ttgttggcat ctctgctgtc ttctgggttg gaagcaaaaa
     2401 gacatgcaca gaatgggctg ggttttttaa acgaaatcgc aagagagatc caatcagtga
     2461 aagtcgaaga gtactacagg aatcatgtga gtttttctta aagcacaatt ctaaagttaa
     2521 acacaaaaag aagcactata aaccaagttc acacaagctg aaggtcattt ccaaatccat
     2581 gggaaccagc acaggagcta cagcaaatca tggcacttct gcagtagcaa ttactagcca
     2641 tgattaccta ggacaagaaa ctttgacaga aatccaaacc tcaccagaaa catcaatgag
     2701 agaggtgaaa gcggacggag ctagcacccc caggttaaga gaacaggact gtggtgaacc
     2761 tgcctcgcca gcagcatcca tctccagact ctctggggaa caggtcgacg ggaagggcca
     2821 ggcaggcagt gtatctgaaa gtgcgcggag tgaaggaagg attagtccaa agagtgatat
     2881 tactgacact ggcctggcac agagcaacaa tttgcaggtc cccagttctt cagaaccaag
     2941 cagcctcaaa ggttccacat ctctgcttgt tcacccggtt tcaggagtga gaaaagagca
     3001 gggaggtggt tgtcattcag atactctcga ggaacaaaaa ctaatatcag aggaagattt
     3061 ggaacagaaa ctaatcagtg aagaagattt agagcaaaag ttaatttccg aggaggactt
     3121 atgagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3181 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3241 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3301 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3361 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3421 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3481 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3541 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3601 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3661 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3721 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3781 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3841 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3901 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3961 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4021 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4081 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4141 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4201 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4261 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4321 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4381 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4441 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4501 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4561 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4621 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4681 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4741 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4801 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4861 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4921 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4981 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5041 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5101 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5161 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5221 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5281 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5341 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5401 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5461 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5521 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5581 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5641 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5701 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5761 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5821 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5881 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5941 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6001 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6061 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6121 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6181 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6241 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6301 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     6361 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6421 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6481 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6541 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6601 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6661 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6721 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6781 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6841 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6901 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6961 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7021 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7081 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7141 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7201 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7261 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7321 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7381 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7441 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7501 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     7561 gatcgggaga tctcccgatc ccctatggt
//
