LOCUS       V020333                 7553 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020333
VERSION     V020333
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..2989
                     /label="FZD8(NM_031866)"
                     /note="FZD8(NM_031866)"
                     /gene="FZD8"
     CDS             2996..3085
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    3120..3344
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3390..3818
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3832..4161
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4228..5022
                     /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    5196..5317
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5366..5382)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5390..5406
                     /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(5414..5444)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5459..5480
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5768..6353)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6524..7384)
                     /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(7385..7489)
                     /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 gagtggggtt acctgttgga agtgacctcg ctgctggccg ccttggcgct
      961 gctgcagcgc tctagcggcg ctgcggccgc ctcggccaag gagctggcat gccaagagat
     1021 caccgtgccg ctgtgtaagg gcatcggcta caactacacc tacatgccca atcagttcaa
     1081 ccacgacacg caagacgagg cgggcctgga ggtgcaccag ttctggccgc tggtggagat
     1141 ccagtgctcg cccgatctca agttcttcct gtgcagcatg tacacgccca tctgcctaga
     1201 ggactacaag aagccgctgc cgccctgccg ctcggtgtgc gagcgcgcca aggccggctg
     1261 cgcgccgctc atgcgccagt acggcttcgc ctggcccgac cgcatgcgct gcgaccggct
     1321 gcccgagcaa ggcaaccctg acacgctgtg catggactac aaccgcaccg acctaaccac
     1381 cgccgcgccc agcccgccgc gccgcctgcc gccgccgccg cccggcgagc agccgccttc
     1441 gggcagcggc cacggccgcc cgccgggggc caggcccccg caccgcggcg gcggcagggg
     1501 cggtggcggc ggggacgcgg cggcgccccc agctcgcggc ggcggcggtg gcgggaaggc
     1561 gcggccccct ggcggcggcg cggctccctg cgagcccggg tgccagtgcc gcgcgcctat
     1621 ggtgagcgtg tccagcgagc gccacccgct ctacaaccgc gtcaagacag gccagatcgc
     1681 taactgcgcg ctgccctgcc acaacccctt tttcagccag gacgagcgcg ccttcaccgt
     1741 cttctggatc ggcctgtggt cggtgctctg cttcgtgtcc accttcgcca ccgtctccac
     1801 cttccttatc gacatggagc gcttcaagta cccggagcgg cccattatct tcctctcggc
     1861 ctgctacctc ttcgtgtcgg tgggctacct agtgcgcctg gtggcgggcc acgagaaggt
     1921 ggcgtgcagc ggtggcgcgc cgggcgcggg gggcgctggg ggcgcgggcg gcgcggcggc
     1981 gggcgcgggc gcggcgggcg cgggcgcggg cggcccgggc gggcgcggcg agtacgagga
     2041 gctgggcgcg gtggagcagc acgtgcgcta cgagaccacc ggccccgcgc tgtgcaccgt
     2101 ggtcttcttg ctggtctact tcttcggcat ggccagctcc atctggtggg tgatcttgtc
     2161 gctcacatgg ttcctggcgg ccggtatgaa gtggggcaac gaagccatcg ccggctactc
     2221 gcagtacttc cacctggccg cgtggcttgt gcccagcgtc aagtccatcg cggtgctggc
     2281 gctcagctcg gtggacggcg acccggtggc gggcatctgc tacgtgggca accagagcct
     2341 ggacaacctg cgcggcttcg tgctggcgcc gctggtcatc tacctcttca tcggcaccat
     2401 gttcctgctg gccggcttcg tgtccctctt ccgcatccgc tcggtcatca agcaacagga
     2461 cggccccacc aagacgcaca agctggagaa gctgatgatc cgcctgggcc tgttcaccgt
     2521 gctctacacc gtgcccgccg cggtggtggt cgcctgcctc ttctacgagc agcacaaccg
     2581 cccgcgctgg gaggccacgc acaactgccc gtgcctgcgg gacctgcagc ccgaccaggc
     2641 acgcaggccc gactacgccg tcttcatgct caagtacttc atgtgcctag tggtgggcat
     2701 cacctcgggc gtgtgggtct ggtccggcaa gacgctggag tcctggcgct ccctgtgcac
     2761 ccgctgctgc tgggccagca agggcgccgc ggtgggcggg ggcgcgggcg ccacggccgc
     2821 ggggggtggc ggcgggccgg ggggcggcgg cggcggggga cccggcggcg gcggggggcc
     2881 gggcggcggc gggggctccc tctacagcga cgtcagcact ggcctgacgt ggcggtcggg
     2941 cacggcgagc tccgtgtctt atccaaagca gatgccattg tcccaggtcc tcgaggaaca
     3001 aaaactaata tcagaggaag atttggaaca gaaactaatc agtgaagaag atttagagca
     3061 aaagttaatt tccgaggagg acttatgagg gcccgtttaa acccgctgat cagcctcgac
     3121 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3181 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3241 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3301 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3361 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3421 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3481 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3541 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3601 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3661 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3721 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3781 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3841 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3901 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3961 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4021 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4081 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4141 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4201 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4261 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4321 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4381 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4441 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4501 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4561 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4621 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4681 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4741 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4801 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4861 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4921 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4981 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5041 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5101 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5161 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5221 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5281 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5341 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5401 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5461 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5521 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5581 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5641 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5701 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5761 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5821 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5881 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5941 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6001 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6061 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6121 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6181 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6241 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6301 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6361 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6421 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6481 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6541 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6601 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6661 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6721 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6781 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6841 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6901 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6961 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7021 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7081 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7141 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7201 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7261 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7321 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7381 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7441 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7501 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
