LOCUS       V026742                 7594 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026742
VERSION     V026742
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             997..3114
                     /label="FZD6(NM_003506)"
                     /note="FZD6(NM_003506)"
                     /gene="FZD6"
     polyA_signal    3194..3418
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3464..3892
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3906..4235
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4302..5096
                     /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    5270..5391
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5440..5456)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5464..5480
                     /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(5488..5518)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5533..5554
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5842..6427)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6598..7458)
                     /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(7459..7563)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt ggtaccatgg aaatgtttac atttttgttg
     1021 acgtgtattt ttctacccct cctaagaggg cacagtctct tcacctgtga accaattact
     1081 gttcccagat gtatgaaaat ggcctacaac atgacgtttt tccctaatct gatgggtcat
     1141 tatgaccaga gtattgccgc ggtggaaatg gagcattttc ttcctctcgc aaatctggaa
     1201 tgttcaccaa acattgaaac tttcctctgc aaagcatttg taccaacctg catagaacaa
     1261 attcatgtgg ttccaccttg tcgtaaactt tgtgagaaag tatattctga ttgcaaaaaa
     1321 ttaattgaca cttttgggat ccgatggcct gaggagcttg aatgtgacag attacaatac
     1381 tgtgatgaga ctgttcctgt aacttttgat ccacacacag aatttcttgg tcctcagaag
     1441 aaaacagaac aagtccaaag agacattgga ttttggtgtc caaggcatct taagacttct
     1501 gggggacaag gatataagtt tctgggaatt gaccagtgtg cgcctccatg ccccaacatg
     1561 tattttaaaa gtgatgagct agagtttgca aaaagtttta ttggaacagt ttcaatattt
     1621 tgtctttgtg caactctgtt cacattcctt acttttttaa ttgatgttag aagattcaga
     1681 tacccagaga gaccaattat atattactct gtctgttaca gcattgtatc tcttatgtac
     1741 ttcattggat ttttgctagg cgatagcaca gcctgcaata aggcagatga gaagctagaa
     1801 cttggtgaca ctgttgtcct aggctctcaa aataaggctt gcaccgtttt gttcatgctt
     1861 ttgtattttt tcacaatggc tggcactgtg tggtgggtga ttcttaccat tacttggttc
     1921 ttagctgcag gaagaaaatg gagttgtgaa gccatcgagc aaaaagcagt gtggtttcat
     1981 gctgttgcat ggggaacacc aggtttcctg actgttatgc ttcttgctat gaacaaagtt
     2041 gaaggagaca acattagtgg agtttgcttt gttggccttt atgacctgga tgcttctcgc
     2101 tactttgtac tcttgccact gtgcctttgt gtgtttgttg ggctctctct tcttttagct
     2161 ggcattattt ccttaaatca tgttcgacaa gtcatacaac atgatggccg gaaccaagaa
     2221 aaactaaaga aatttatgat tcgaattgga gtcttcagcg gcttgtatct tgtgccatta
     2281 gtgacacttc tcggatgtta cgtctatgag caagtgaaca ggattacctg ggagataact
     2341 tgggtctctg atcattgtcg tcagtaccat atcccatgtc cttatcaggc aaaagcaaaa
     2401 gctcgaccag aattggcttt atttatgata aaatacctga tgacattaat tgttggcatc
     2461 tctgctgtct tctgggttgg aagcaaaaag acatgcacag aatgggctgg gttttttaaa
     2521 cgaaatcgca agagagatcc aatcagtgaa agtcgaagag tactacagga atcatgtgag
     2581 tttttcttaa agcacaattc taaagttaaa cacaaaaaga agcactataa accaagttca
     2641 cacaagctga aggtcatttc caaatccatg ggaaccagca caggagctac agcaaatcat
     2701 ggcacttctg cagtagcaat tactagccat gattacctag gacaagaaac tttgacagaa
     2761 atccaaacct caccagaaac atcaatgaga gaggtgaaag cggacggagc tagcaccccc
     2821 aggttaagag aacaggactg tggtgaacct gcctcgccag cagcatccat ctccagactc
     2881 tctggggaac aggtcgacgg gaagggccag gcaggcagtg tatctgaaag tgcgcggagt
     2941 gaaggaagga ttagtccaaa gagtgatatt actgacactg gcctggcaca gagcaacaat
     3001 ttgcaggtcc ccagttcttc agaaccaagc agcctcaaag gttccacatc tctgcttgtt
     3061 cacccggttt caggagtgag aaaagagcag ggaggtggtt gtcattcaga tacttgagaa
     3121 ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc
     3181 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3241 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3301 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3361 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3421 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3481 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3541 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3601 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3661 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3721 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3781 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3841 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3901 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3961 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4021 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4081 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4141 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4201 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4261 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4321 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4381 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4441 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4501 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4561 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4621 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4681 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4741 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4801 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4861 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4921 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4981 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5041 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5101 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5161 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5221 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5281 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5341 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5401 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5461 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5521 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5581 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5641 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5701 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5761 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5821 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5881 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5941 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6001 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6061 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6121 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6181 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6241 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6301 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6361 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6421 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6481 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6541 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6601 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6661 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6721 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6781 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6841 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6901 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6961 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7021 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7081 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7141 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7201 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7261 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7321 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7381 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7441 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7501 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7561 gcgcacattt ccccgaaaag tgccacctga cgtc
//
