LOCUS       V030267                 8257 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030267
VERSION     V030267
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"
     CDS             706..2913
                     /label="Dvl2(NM_001172056)"
                     /note="Dvl2(NM_001172056)"
                     /gene="Dvl2"
     CDS             2920..3009
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3033..3621
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3687..3735
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3937..4365
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4379..4708
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4775..5569
                     /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    5745..5866
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5915..5931)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5939..5955
                     /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(5963..5993)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6008..6029
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6317..6905)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7076..7936)
                     /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(7937..8041)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc gggcagcagc
      721 gcggggggcg gtggtgtagg cgagacgaag gtgatttacc atctggatga agaagagact
      781 ccttacctgg tgaagatccc tgtcccggcg gagcgcatca cgctcggcga tttcaagagc
      841 gttttgcagc ggcccgcggg cgctaagtac tttttcaagt ctatggatca ggattttggg
      901 gtggtgaagg aagagatctc tgacgacagt gcccgcctac cttgcttcaa tggaagggtt
      961 gtctcctggc tggtgtcatc agagagcccc cagcctgaga cagcgccccc tgcccatgag
     1021 tctcggacag aactggttcc tccacctcca ccattacccc ctttgccacc agaaaggacc
     1081 agtggcattg gggactcaag gcctccatcc ttccacccta atgtatccag tagccatgaa
     1141 aatctagagc ctgagacaga aactgaatcc gtcgtatcac tgagacgaga ccgacctagg
     1201 aggagagaca gcagtgagca tggcgctggt ggccacaggc ctggcgtccc ctcaaggctg
     1261 gagcgccacc tggctgggta tgagagctct tccaccctca tgaccagcga actggaaagt
     1321 accagcttag gagattcaga tgaggatgac agcatgagta ggttcagcag ctccactgag
     1381 cagagcagtg cctcccgcct cctcaagcgc caccgaaggc gaaggaagca gcggccgcca
     1441 cgcatggaga ggacctcatc cttcagcagt gtcacagatt ctacaatgtc tctcaacatc
     1501 atcacagtca cactcaacat ggagaaatac aacttcctgg gtatctccat tgtgggccaa
     1561 agtaacgagc gtggtgacgg aggcatctat ataggctcca tcatgaaagg gggagcggtg
     1621 gctgcagatg ggcggatcga gcctggagac atgcttttgc aggtaaacga catgaacttt
     1681 gagaacatga gcaatgacga tgctgtgcgg gtgctgagag acattgtaca caagccaggc
     1741 cccatcgtgc tcaccgtggc caagtgttgg gatccctctc cccaggccta cttcaccctc
     1801 ccccgaaatg agcccataca gccaattgat ccagctgcct gggtgtccca ttcagctgcg
     1861 ctaaccggtg ccttccctgc ttaccccggc tcttcatcta tgagcactat cacatctggc
     1921 tcctctctgc ctgatggctg tgaaggccgg ggtctctctg tccacatgga catggcctct
     1981 gtgaccaagg ccatggcagc cccagagtct gggcttgaag tccgggaccg catgtggctc
     2041 aagatcacca tcccaaacgc ctttctaggc tcggatgtgg tagactggct gtatcaccat
     2101 gtggaaggct ttcctgagcg ccgggaggcc cgcaagtacg ccagtgggct gttgaaggcg
     2161 ggactcatcc ggcacactgt caacaagatt actttctctg agcagtgcta ctatgttttc
     2221 ggggacctca gtggtggctg cgagagttac ctagttaacc tgtctctgaa tgacaacgat
     2281 ggttccagtg gggcttcaga ccaggatact ctggctcctc ttcctggagc tactccctgg
     2341 cccctgctgc ctaccttctc ctaccaatac ccagccccgc acccctacag cccacagcct
     2401 ccgccctatc atgagctttc gtcctacagc tatggcggag gcagtgccag cagccagcac
     2461 agtgagggga gccggagcag tgggtcaaca agaagtgatg ggggggccgg gcgcacaggg
     2521 aggcctgagg aacgggcccc tgagtcaaag tctggcagtg gcagtgagtc tgaactctcc
     2581 agccggggag gcagccttcg gcggggtggg gaacctggtg ggactggtga tggtggaccc
     2641 cctccatcca ggggctcagc aggtggtact cctaacctcc gagctctccc aggcctccat
     2701 ccctatggac ctccacctgg catggctctc ccctataacc ccatgatggt agtcatgatg
     2761 cctccacccc caccccctgt ctcgacagca gtacagcccc ctggtgcccc tccagtcaga
     2821 gacctgggct ctgtgccccc agaactgaca gctagccgtc agagcttcca catggccatg
     2881 ggcaacccca gtgagttttt tgtagatgtt atgctcgagg aacaaaaact aatatcagag
     2941 gaagatttgg aacagaaact aatcagtgaa gaagatttag agcaaaagtt aatttccgag
     3001 gaggacttat agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg
     3061 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3121 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3181 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3241 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3301 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3361 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3421 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     3481 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3541 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3601 ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac
     3661 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg
     3721 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     3781 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     3841 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     3901 cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3961 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4021 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4081 ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4141 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4201 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4261 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct
     4321 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4381 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4441 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4501 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4561 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4621 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4681 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4741 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4801 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4861 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4921 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4981 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5041 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5101 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5161 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5221 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5281 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5341 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5401 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5461 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5521 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5581 gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5641 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5701 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5761 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5821 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5881 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5941 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6001 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6061 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6121 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6181 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6241 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6301 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6361 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6421 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6481 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     6541 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6601 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6661 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6721 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6781 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6841 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6901 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6961 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7021 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7081 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7141 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7201 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7261 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7321 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7381 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7441 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7501 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7561 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7621 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7681 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7741 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7801 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7861 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7921 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7981 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8041 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     8101 atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     8161 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     8221 aggcttgacc gacaattgca tgaagaatct gcttagg
//
