LOCUS       V029177                 7621 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029177
VERSION     V029177
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"
     CDS             964..3111
                     /label="Dvl3(NM_007889)"
                     /note="Dvl3(NM_007889)"
                     /gene="Dvl3"
     CDS             3118..3183
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3221..3445
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3491..3919
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3933..4262
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4329..5123
                     /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    5297..5418
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5467..5483)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5491..5507
                     /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(5515..5545)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5560..5581
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5869..6454)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6625..7485)
                     /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(7486..7590)
                     /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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgggcg agaccaagat catctaccac ctggacggcc aggagacgcc gtacctggtg
     1021 aagctgcccc tgcccgccga gcgcgtcacc ttggcggact ttaagggcgt tctgcagcgg
     1081 cccagctata agttcttctt caagtctatg gacgacgatt ttggggtggt gaaggaagag
     1141 atctcggacg acaatgccaa gctaccctgc ttcaatggcc gggtggtgtc ctggctggta
     1201 tctgccgagg gctcacaccc agagccggcc cctttctgtg ctgacaaccc gtcagagctg
     1261 ccaccttcga tggaacgcac aggaggcatt ggggactctc gacccccctc cttccaccct
     1321 catgctagtg ggggcagcca ggagaatctg gacaatgaca cggagaccga ctccttggtg
     1381 tctgcccaac gagagcggcc ccgacggagg gatggcccgg agcacgcagc ccgactcaat
     1441 ggaaccacaa agggtgagcg gcgacgggag ccaggcggct atgacagttc atctaccctc
     1501 atgagcagcg agctggagac taccagtttc tttgattccg atgaggatga ttccaccagc
     1561 aggttcagca gctccacaga acagagcagt gcctcacgcc tcatgagaag acacaagcgg
     1621 cggcggcgga agcagaaggt ctcccggatt gagcggtcct cgtctttcag cagcatcaca
     1681 gactccacta tgtctctcaa catcatcacc gtcactctca acatggaaaa gtataacttc
     1741 ttgggcatct ccattgtggg ccaaagcaat gagcgaggtg atgggggcat ctacattggc
     1801 tccatcatga agggtggggc tgtggcagct gatggacgca ttgagccagg agatatgcta
     1861 ctgcaggtaa atgagatcaa cttcgagaac atgagcaacg atgacgcagt ccgagtcctt
     1921 cgggagattg tgcacaagcc agggcccatc actctgactg tagccaagtg ctgggaccca
     1981 agtcctcgcg gctgcttcac attaccccgg agcgagccca tccgtcccat tgaccctgca
     2041 gcctgggtct ctcacacggc agccatgaca ggcaccttcc ccgcctatgg catgagcccc
     2101 tccttaagca ccatcacctc taccagctcc tccatcacca gctccatccc tgatacagag
     2161 cgcctagatg atttccacct ctccatccac agtgacatgg cggccatcgt aaaagccatg
     2221 gcctcccctg agtcagggtt ggaggtccga gaccgcatgt ggctcaagat taccattccc
     2281 aatgctttca tcggctcgga tgtggtggac tggctgtacc acaacgtgga agggttcacc
     2341 gaccgtcggg aggcccgcaa gtatgctagc aacctgctga aagctggttt catccgccat
     2401 accgtcaaca agatcacgtt ctccgagcag tgctactaca tctttggcga cctctgtggt
     2461 aacatggcca acctatcgct tcacgaccat gatggctcca gtggcgcctc tgaccaggac
     2521 acactggctc ctttgccgca cccaggagct gccccttggc ctatggcttt cccttaccag
     2581 tacccaccac ctccgcatcc ctacaaccca cacccaggct tcccagaact gggctacagc
     2641 tacggtgggg gcagcgccag cagtcagcac agtgaaggca gtcggagcag tggctccaac
     2701 cgaagtggca gcgaccggcg caaggagaag gacccaaaag ctggggactc caagtcaggt
     2761 ggtagcggca gcgagtcaga ccacactaca cgcagcagtc tgagggggcc acgggagcgg
     2821 gcaccaagcg agcgatcagg cccagcggcc agcgagcaca gccaccgcag tcaccattcg
     2881 ctgaccagca gcctgcgcag ccaccacaca cacccaagct atggcccacc cggagtgccc
     2941 cctctctatg gtcccccgat gctgatgatg accccgccgc cggcagccat gggtccccca
     3001 ggagcccctc cagggcgtga cctggcctcg gtgcccccgg aactgacggc cagcagacag
     3061 tccttccgca tggcaatggg aaaccccagt gagttctttg tggatgtgat gctcgaggac
     3121 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     3181 aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     3241 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3301 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3361 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3421 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3481 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3541 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3601 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3661 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3721 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3781 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3841 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3901 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3961 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4021 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4081 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4141 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4201 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4261 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4321 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4381 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4441 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4501 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4561 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4621 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4681 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4741 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4801 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4861 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4921 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4981 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5041 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5101 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5161 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5221 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5281 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5341 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5401 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5461 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5521 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5581 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5641 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5701 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5761 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5821 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5881 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5941 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6001 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6061 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6121 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6181 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6241 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6301 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6361 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa
     6421 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6481 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6541 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6601 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6661 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6721 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6781 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6841 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6901 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6961 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7021 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7081 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7141 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7201 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7261 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7321 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7381 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7441 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7501 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7561 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7621 c
//
