LOCUS       V022151                 9168 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022151
VERSION     V022151
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        431..708
                     /label="chicken β-actin promoter"
     intron          709..1726
                     /note="chimera between introns from chicken β-actin and
                     rabbit β-globin"
                     /label="chimeric intron"
     CDS             1839..3836
                     /label="FZD3(NM_017412)"
                     /note="FZD3(NM_017412)"
                     /gene="FZD3"
     CDS             3843..3932
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3944..4532
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4598..4646
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4848..5276
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5290..5619
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5686..6480
                     /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    6656..6777
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6826..6842)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6850..6866
                     /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(6874..6904)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6919..6940
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7228..7816)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7987..8847)
                     /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(8848..8952)
                     /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 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
      481 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
      541 gggggggggg ggcccccccc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
      601 cggaaaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
      661 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgttgcg
      721 cgctgccttc cccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
      781 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      841 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
      901 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc
      961 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc
     1021 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg
     1081 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg
     1141 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
     1201 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
     1261 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
     1321 gccggggagg gctcggggga aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg
     1381 cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc
     1441 tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg
     1501 gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc
     1561 gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg
     1621 ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc
     1681 tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt
     1741 gctggttatt gtgctgtctc atcattttgg caaagaattc tgcagctggc tagcgtttaa
     1801 acttaagctt ggtaccgagc tcggatccag atgccaccat ggctatgact tggattgtct
     1861 tctctctttg gcccttgact gtgttcatgg ggcatatagg tgggcacagt ttgttttctt
     1921 gtgaacctat taccttgagg atgtgccaag atttgcctta taatactacc ttcatgccta
     1981 atcttctgaa tcattatgac caacagacag cagctttggc aatggagcca ttccacccta
     2041 tggtgaatct ggattgttct cgggatttcc ggccttttct ttgtgcactc tacgctccta
     2101 tttgtatgga atatggacgt gtcacacttc cctgtcgtag gctgtgtcag cgggcttaca
     2161 gtgagtgttc gaagctcatg gagatgtttg gtgttccttg gcctgaagat atggaatgca
     2221 gtaggttccc agattgtgat gagccatatc ctcgacttgt ggatctgaat ttagctggag
     2281 aaccaactga aggagcccca gtggcagtgc agagagacta tggtttttgg tgtccccgag
     2341 agttaaaaat tgatcctgat ctgggttatt cttttctgca tgtgcgtgat tgttcacctc
     2401 cttgtccaaa tatgtacttc agaagagaag aactgtcatt tgctcgctat ttcataggat
     2461 tgatttcaat catttgcctc tcggccacat tgtttacttt tttaactttt ttgattgatg
     2521 tcacaagatt ccgttatcct gaaaggccta ttatatttta tgcagtctgc tacatgatgg
     2581 tatccttaat tttcttcatt ggatttttgc ttgaagatcg agtagcctgc aatgcatcca
     2641 tccctgcaca atataaggct tccacagtga cacaaggatc tcataataaa gcctgtacca
     2701 tgctttttat gatactctat ttttttacta tggctggcag tgtatggtgg gtaattctta
     2761 ccatcacatg gtttttagca gctgtgccaa agtggggtag tgaagctatt gagaagaaag
     2821 cattgctgtt tcacgccagt gcatggggca tccccggaac tctaaccatc atccttttag
     2881 cgatgaataa aattgaaggt gacaatatta gtggcgtgtg ttttgttggc ctctacgatg
     2941 ttgatgcatt gagatatttt gttcttgctc ccctctgcct gtatgtggta gttggggttt
     3001 ctctcctctt agctggcatt atatccctaa acagagttcg aattgagatt ccattagaaa
     3061 aggagaacca agataaatta gtgaagttta tgatccggat cggtgttttc agcattcttt
     3121 atctcgtacc actcttggtt gtaattggat gctactttta tgagcaagct taccggggca
     3181 tctgggaaac aacgtggata caagaacgct gcagagaata tcacattcca tgtccatatc
     3241 aggttactca aatgagtcgt ccagacttga ttctctttct gatgaaatac ctgatggctc
     3301 tcatagttgg cattccctct gtattttggg ttggaagcaa aaagacatgc tttgaatggg
     3361 ccagtttttt tcatggtcgt aggaaaaaag agatagtgaa tgagagccga caggtactcc
     3421 aggaacctga ttttgctcag tctctcctga gggatccaaa tactcctatc ataagaaagt
     3481 caaggggaac ttccactcaa ggaacatcca cccatgcttc ttcaactcag ctggctatgg
     3541 tggatgatca aagaagcaaa gcaggaagca tccacagcaa agtgagcagc taccacggca
     3601 gcctccacag atcacgtgat ggcaggtaca cgccctgcag ttacagagga atggaggaga
     3661 gactacctca tggcagcatg tcacgactaa cagatcactc caggcatagt agttctcatc
     3721 ggctcaatga acagtcacga catagcagca tcagagatct cagtaataat cccatgactc
     3781 atatcacaca tggcaccagc atgaatcggg ttattgaaga agatggaacc agtgctctcg
     3841 aggaacaaaa actaatatca gaggaagatt tggaacagaa actaatcagt gaagaagatt
     3901 tagagcaaaa gttaatttcc gaggaggact tatagatctc gacaatcaac ctctggatta
     3961 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     4021 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     4081 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     4141 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     4201 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     4261 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     4321 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     4381 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     4441 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     4501 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     4561 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4621 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     4681 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     4741 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     4801 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     4861 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4921 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4981 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     5041 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5101 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5161 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     5221 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     5281 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     5341 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5401 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5461 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5521 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     5581 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     5641 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5701 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5761 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5821 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     5881 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5941 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6001 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6061 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6121 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6181 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     6241 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6301 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6361 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6421 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6481 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     6541 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6601 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     6661 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6721 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6781 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     6841 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6901 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6961 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7021 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7081 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7141 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7201 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7261 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7321 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7381 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     7441 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7501 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7561 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7621 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7681 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7741 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7801 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7861 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7921 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7981 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8041 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8101 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8161 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8221 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8281 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8341 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8401 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8461 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8521 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8581 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8641 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8701 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8761 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8821 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8881 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8941 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     9001 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     9061 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     9121 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
