LOCUS       V033923                 8193 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033923
VERSION     V033923
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2873
                     /label="FZD6(NM_003506)"
                     /note="FZD6(NM_003506)"
                     /gene="FZD6"
     CDS             2880..2945
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2969..3557
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3623..3671
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3873..4301
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4315..4644
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4711..5505
                     /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    5681..5802
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5851..5867)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5875..5891
                     /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(5899..5929)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5944..5965
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6253..6841)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7012..7872)
                     /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(7873..7977)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga aatgtttaca tttttgttga
      781 cgtgtatttt tctacccctc ctaagagggc acagtctctt cacctgtgaa ccaattactg
      841 ttcccagatg tatgaaaatg gcctacaaca tgacgttttt ccctaatctg atgggtcatt
      901 atgaccagag tattgccgcg gtggaaatgg agcattttct tcctctcgca aatctggaat
      961 gttcaccaaa cattgaaact ttcctctgca aagcatttgt accaacctgc atagaacaaa
     1021 ttcatgtggt tccaccttgt cgtaaacttt gtgagaaagt atattctgat tgcaaaaaat
     1081 taattgacac ttttgggatc cgatggcctg aggagcttga atgtgacaga ttacaatact
     1141 gtgatgagac tgttcctgta acttttgatc cacacacaga atttcttggt cctcagaaga
     1201 aaacagaaca agtccaaaga gacattggat tttggtgtcc aaggcatctt aagacttctg
     1261 ggggacaagg atataagttt ctgggaattg accagtgtgc gcctccatgc cccaacatgt
     1321 attttaaaag tgatgagcta gagtttgcaa aaagttttat tggaacagtt tcaatatttt
     1381 gtctttgtgc aactctgttc acattcctta cttttttaat tgatgttaga agattcagat
     1441 acccagagag accaattata tattactctg tctgttacag cattgtatct cttatgtact
     1501 tcattggatt tttgctaggc gatagcacag cctgcaataa ggcagatgag aagctagaac
     1561 ttggtgacac tgttgtccta ggctctcaaa ataaggcttg caccgttttg ttcatgcttt
     1621 tgtatttttt cacaatggct ggcactgtgt ggtgggtgat tcttaccatt acttggttct
     1681 tagctgcagg aagaaaatgg agttgtgaag ccatcgagca aaaagcagtg tggtttcatg
     1741 ctgttgcatg gggaacacca ggtttcctga ctgttatgct tcttgctatg aacaaagttg
     1801 aaggagacaa cattagtgga gtttgctttg ttggccttta tgacctggat gcttctcgct
     1861 actttgtact cttgccactg tgcctttgtg tgtttgttgg gctctctctt cttttagctg
     1921 gcattatttc cttaaatcat gttcgacaag tcatacaaca tgatggccgg aaccaagaaa
     1981 aactaaagaa atttatgatt cgaattggag tcttcagcgg cttgtatctt gtgccattag
     2041 tgacacttct cggatgttac gtctatgagc aagtgaacag gattacctgg gagataactt
     2101 gggtctctga tcattgtcgt cagtaccata tcccatgtcc ttatcaggca aaagcaaaag
     2161 ctcgaccaga attggcttta tttatgataa aatacctgat gacattaatt gttggcatct
     2221 ctgctgtctt ctgggttgga agcaaaaaga catgcacaga atgggctggg ttttttaaac
     2281 gaaatcgcaa gagagatcca atcagtgaaa gtcgaagagt actacaggaa tcatgtgagt
     2341 ttttcttaaa gcacaattct aaagttaaac acaaaaagaa gcactataaa ccaagttcac
     2401 acaagctgaa ggtcatttcc aaatccatgg gaaccagcac aggagctaca gcaaatcatg
     2461 gcacttctgc agtagcaatt actagccatg attacctagg acaagaaact ttgacagaaa
     2521 tccaaacctc accagaaaca tcaatgagag aggtgaaagc ggacggagct agcaccccca
     2581 ggttaagaga acaggactgt ggtgaacctg cctcgccagc agcatccatc tccagactct
     2641 ctggggaaca ggtcgacggg aagggccagg caggcagtgt atctgaaagt gcgcggagtg
     2701 aaggaaggat tagtccaaag agtgatatta ctgacactgg cctggcacag agcaacaatt
     2761 tgcaggtccc cagttcttca gaaccaagca gcctcaaagg ttccacatct ctgcttgttc
     2821 acccggtttc aggagtgaga aaagagcagg gaggtggttg tcattcagat actggatccg
     2881 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     2941 acaagtagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     3001 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3061 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3121 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3181 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3241 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3301 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3361 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3421 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3481 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3541 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     3601 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     3661 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3721 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     3781 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     3841 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3901 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3961 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4021 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4081 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4141 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4201 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     4261 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4321 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4381 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4441 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4501 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4561 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4621 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4681 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4741 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4801 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4861 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4921 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4981 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5041 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5101 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5161 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5221 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5281 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5341 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5401 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5461 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5521 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5581 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5641 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5701 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5761 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5821 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5881 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5941 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6001 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6061 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6121 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6181 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6241 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6301 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6361 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6421 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     6481 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6541 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6601 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6661 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6721 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6781 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6841 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6901 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6961 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7021 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7081 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7141 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7201 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7261 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7321 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7381 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7441 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7501 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7561 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7621 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7681 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7741 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7801 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7861 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7921 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7981 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     8041 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     8101 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     8161 ttgaccgaca attgcatgaa gaatctgctt agg
//
