LOCUS       V028690                 8607 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028690
VERSION     V028690
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3287
                     /label="PRICKLE2(NM_001370528)"
                     /note="PRICKLE2(NM_001370528)"
                     /gene="PRICKLE2"
     CDS             3294..3359
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3383..3971
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4037..4085
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4287..4715
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4729..5058
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5125..5919
                     /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    6095..6216
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6265..6281)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6289..6305
                     /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(6313..6343)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6358..6379
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6667..7255)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7426..8286)
                     /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(8287..8391)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggt gacagtgatg ccgctggaga
      781 tggagaagac catcagcaaa ctcatgtttg actttcagag gaactcgacc tcagatgatg
      841 actcaggctg tgctttggaa gagtatgcct gggtcccgcc gggtctgaag cctgaacagg
      901 tacaccagta ctatagctgt ctcccagaag agaaagtccc ttatgtcaac agtcctggag
      961 agaaactgcg aatcaagcag ctactacacc agctgccgcc acatgacaat gaggttcgat
     1021 attgcaactc cctggatgag gaagagaaga gggagctgaa gcttttcagc agccagagga
     1081 aacgcgaaaa cttgggccgc gggaatgtca ggcctttccc agtcaccatg acaggagcta
     1141 tttgtgaaca gtgcggaggc cagatcaatg gtggagacat cgctgtgttt gcgtcacgcg
     1201 ctggccacgg cgtttgctgg cacccgccgt gcttcgtatg cactgtctgc aatgagctcc
     1261 tggtggatct gatctacttt taccaagatg ggaagatata ctgtggcagg caccatgctg
     1321 agtgcctgaa gccgcgctgt gctgcctgcg atgagatcat ctttgcagat gaatgcacag
     1381 aagctgaggg gcgacactgg cacatgaaac acttttgctg cttcgagtgt gagacagtgc
     1441 tgggcggcca gcgctacatc atgaaggagg gaagacccta ctgttgccac tgcttcgagt
     1501 ccttgtatgc agaatattgt gacacctgtg cccaacatat aggtatcgac caaggtcaaa
     1561 tgacctatga tggccaacac tggcatgcca ctgagacctg tttctgctgt gctcactgca
     1621 agaaatccct cctggggcgg ccattcctcc cgaagcaggg ccagatattc tgctcacggg
     1681 cctgcagtgc tggggaagac cccaatggtt ctgactcctc tgattccgcc ttccagaacg
     1741 ccagggccaa ggagtcccgg cgcagtgcca aaattggcaa gaacaagggc aagacggagg
     1801 agcccatgct gaaccagcac agccagctgc aagtgagttc taaccggctg tcagccgacg
     1861 tagaccccct gtcactgcag atggacatgc tcagcctgtc cagccagaca cccagcctca
     1921 accgggaccc catctggagg agccgggaag agccctacca ttatgggaac aagatggagc
     1981 agaaccagac ccagagccct ctgcagctcc tcagccagtg caacatcaga acttcctaca
     2041 gtccaggagg gcaaggggct ggggcccagc ccgaaatgtg gggcaagcac ttcagcaacc
     2101 ccaaaaggag ctcgtcactg gccatgacag gacatgctgg cagcttcatc aaggaatgcc
     2161 gagaagacta ttacccgggg aggctgagat ctcaggagag ctacagtgat atgtctagtc
     2221 agagtttcag tgagacccga ggcagcatcc aagtccccaa atatgaggag gaagaggaag
     2281 aggaaggggg cttgtccact cagcagtgtc ggacccgtca tcccatcagt tccctgaaat
     2341 acacagagga catgacgccc acagagcaga cccctcgggg ctccatggaa tccctggccc
     2401 tgtctaatgc aacaggcctc tctgctgatg gtggtgccaa gcgccaggag cacctatccc
     2461 gattttccat gcctgacctc agcaaagact ctggaatgaa tgtgtctgag aagctgagca
     2521 acatgggcac tcttaactcg tccatgcagt tccggagcgc agagtcagtt cgcagcctgc
     2581 tctctgccca gcagtaccag gagatggagg gaaacctcca ccagctcagc aaccccattg
     2641 gctacagaga cctgcagtcc cacggaagga tgcatcagag ctttgatttt gatggaggga
     2701 tggcgggcag caagctgcca gggcaggagg gcgtgaggat ccagcccatg agtgaacgca
     2761 cccggagaag agctacttca cgcgacgaca accgccgttt ccgacctcac aggtccaggc
     2821 gttcccgacg ctctcgctcc gacaacgccc tccacctggc cagcgaacgc gaggccatct
     2881 cccggttaaa agataggccc cctctgagag ccagggagga ctatgaccaa tttatgcgcc
     2941 agcggagctt ccaggagagc atggggcatg ggtcccggag ggacctgtac ggccagtgcc
     3001 ctaggactgt gtcggacctg gctttgcaga atgcctttgg ggaccgctgg ggaccctact
     3061 tcgccgagta tgattggtgt tccacctgct cctcctcttc agagtctgac aacgagggct
     3121 atttcctagg agaacccatc ccccagccag cgcgcctgcg atacgtcaca agcgatgagc
     3181 tgctgcacaa atacagctcc tacggcctcc ccaaatcttc cacattaggt ggcagaggac
     3241 agttgcacag caggaaaaga cagaagagca aaaactgtat catttctgga tccgactaca
     3301 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     3361 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     3421 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3481 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3541 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3601 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3661 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3721 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3781 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3841 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3901 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3961 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     4021 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     4081 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4141 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4201 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     4261 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4321 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4381 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4441 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4501 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4561 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4621 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     4681 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4741 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4801 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4861 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4921 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4981 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5041 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     5101 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5161 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5221 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5281 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5341 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5401 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5461 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5521 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5581 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5641 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5701 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5761 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5821 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5881 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     5941 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6001 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6061 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     6121 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6181 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6241 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6301 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6361 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6421 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6481 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6541 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6601 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6661 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6721 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6781 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6841 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6901 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6961 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7021 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7081 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7141 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7201 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7261 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7321 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7381 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7441 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7501 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7561 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7621 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7681 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7741 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7801 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7861 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7921 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7981 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8041 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8101 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8161 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8221 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8281 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8341 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8401 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     8461 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     8521 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     8581 gacaattgca tgaagaatct gcttagg
//
