LOCUS       V026986                 8823 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026986
VERSION     V026986
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..3503
                     /label="CDH4(NM_001794)"
                     /note="CDH4(NM_001794)"
                     /gene="CDH4"
     CDS             3510..3575
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3599..4187
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4253..4301
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4503..4931
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4945..5274
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5341..6135
                     /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    6311..6432
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6481..6497)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6505..6521
                     /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(6529..6559)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6574..6595
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6883..7471)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7642..8502)
                     /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(8503..8607)
                     /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 ccaccatgac cgcgggcgcc ggcgtgctcc
      781 ttctgctgct ctcgctctcc ggcgcgctcc gggcccataa tgaggatctt acaactagag
      841 agacctgcaa ggctgggttc tctgaagatg attacacggc attaatctcc caaaatattc
      901 tagaagggga aaagctactt caagtcaagt tcagcagctg tgtggggacc aaggggacac
      961 aatatgagac caacagcatg gacttcaaag ttggggcaga tgggacagtc ttcgccaccc
     1021 gggagctgca ggtcccctcc gagcaggtgg cgttcacggt gactgcatgg gacagccaga
     1081 cagcagagaa atgggacgcc gtggtgcggt tgctggtggc ccagacctcg tccccgcact
     1141 ctggacacaa gccgcagaaa ggaaagaagg tcgtggctct ggacccctct ccgcctccga
     1201 aggacaccct gctgccgtgg ccccagcacc agaacgccaa cgggctgagg cggcgcaaac
     1261 gggactgggt catcccgccc atcaacgtgc ccgagaactc gcgcgggccc ttcccgcagc
     1321 agctcgtgag gatccggtcc gacaaagaca atgacatccc catccggtac agcatcacgg
     1381 gagtgggcgc cgaccagccc cccatggagg tcttcagcat tgactccatg tccggccgga
     1441 tgtacgtcac aaggcccatg gaccgggagg agcacgcctc ttaccacctc cgagcccacg
     1501 ctgtggacat gaatggcaac aaggtggaga accccatcga cctgtacatc tacgtcatcg
     1561 acatgaatga caaccgccct gagttcatca accaggtcta caacggctcc gtggacgagg
     1621 gctccaagcc aggcacctac gtgatgaccg tcacggccaa cgatgctgac gacagcacca
     1681 cggccaacgg gatggtgcgg taccggatcg tgacccagac cccacagagc ccgtcccaga
     1741 atatgttcac catcaacagc gagactggag atatcgtcac agtggcggct ggcctggacc
     1801 gagagaaagt tcagcagtac acagtcatcg ttcaggccac agatatggaa ggaaatctca
     1861 actatggcct ctcaaacaca gccacagcca tcatcacggt gacagatgtg aatgacaacc
     1921 cgccagaatt taccgccagc acgtttgcag gggaggtccc cgaaaaccgc gtggagaccg
     1981 tggtcgcaaa cctcacggtg atggaccgag atcagcccca ctctccaaac tggaatgccg
     2041 tttaccgcat catcagtggg gatccatccg ggcacttcag cgtccgcaca gaccccgtaa
     2101 ccaacgaggg catggtcacc gtggtgaagg cagtcgacta cgagctcaac agagctttca
     2161 tgctgacagt gatggtgtcc aaccaggcgc ccctggccag cggaatccag atgtccttcc
     2221 agtccacggc aggggtgacc atctccatca tggacatcaa cgaggctccc tacttcccct
     2281 caaaccacaa gctgatccgc ctggaggagg gcgtgccccc cggcaccgtg ctgaccacgt
     2341 tttcagctgt ggaccctgac cggttcatgc agcaggctgt gagatactca aagctgtcag
     2401 acccagcgag ctggctgcac atcaatgcca ccaacggcca gatcaccacg gcggcagtgc
     2461 tggaccgtga gtccctctac accaaaaaca acgtctacga ggccaccttc ctggcagctg
     2521 acaatgggat acccccggcc agcggcaccg ggaccctcca gatctatctc attgacatca
     2581 acgacaacgc ccctgagctg ctgcccaagg aggcgcagat ctgcgagaag cccaacctga
     2641 acgccatcaa catcacggcg gccgacgctg acgtcgaccc caacatcggc ccctacgtct
     2701 tcgagctgcc ctttgtcccg gcggccgtgc ggaagaactg gaccatcacc cgcctgaacg
     2761 gtgactatgc ccaactcagc ttgcgcatcc tgtacctgga ggccgggatg tatgacgtcc
     2821 ccatcatcgt cacagactct ggaaaccctc ccctgtccaa cacgtccatc atcaaagtca
     2881 aggtgtgccc atgtgatgac aacggggact gcaccaccat tggcgcagtg gcagcggctg
     2941 gtctgggcac cggtgccatc gtggccatcc tcatctgcat cctcatcctg ctgaccatgg
     3001 tcctgctgtt tgtcatgtgg atgaagcggc gagagaagga gcgccacacg aagcagctgc
     3061 tcattgaccc cgaggacgac gtccgcgaca acatcctcaa gtatgacgag gaaggcggtg
     3121 gcgaggagga ccaggactac gacctcagcc agctgcagca gccggaagcc atggggcacg
     3181 tgccaagcaa agcccctggc gtgcgtcgcg tggatgagcg gccggtgggc gctgagcccc
     3241 agtacccgat caggcccatg gtgccgcacc caggcgacat cggtgacttc atcaatgagg
     3301 gactccgcgc tgctgacaac gaccccacgg caccccccta tgactccctg ctggtcttcg
     3361 actacgaggg gagcggctcc accgcaggct ccgtcagctc cctgaactca tccagttccg
     3421 gggaccaaga ctacgattac ctcaacgact gggggcccag attcaagaag ctggcggaca
     3481 tgtatggagg tggtgaagag gatggatccg actacaaaga ccatgacggt gattataaag
     3541 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     3601 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3661 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3721 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3781 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3841 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     3901 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3961 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     4021 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     4081 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     4141 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     4201 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4261 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4321 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4381 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4441 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     4501 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4561 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4621 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     4681 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4741 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4801 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4861 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4921 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4981 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5041 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5101 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5161 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5221 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5281 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5341 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5401 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5461 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5521 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5581 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5641 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5701 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5761 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5821 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5881 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5941 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6001 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6061 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6121 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     6181 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6241 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6301 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6361 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6421 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6481 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6541 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6601 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6721 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6781 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     7081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7321 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7561 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7621 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7681 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7741 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7801 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7861 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7921 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7981 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8041 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8101 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8161 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8221 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8281 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8341 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8401 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8461 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8521 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8581 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     8641 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     8701 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     8761 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     8821 agg
//
