LOCUS       V027023                 8466 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027023
VERSION     V027023
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..3192
                     /label="CDH3(NM_001793)"
                     /note="CDH3(NM_001793)"
                     /gene="CDH3"
     CDS             3199..3222
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     misc_feature    3242..3830
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3896..3944
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4146..4574
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4588..4917
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4984..5778
                     /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    5954..6075
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6124..6140)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6148..6164
                     /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(6172..6202)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6217..6238
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6526..7114)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7285..8145)
                     /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(8146..8250)
                     /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 ccaccatggg gctccctcgt
      721 ggacctctcg cgtctctcct ccttctccag gtttgctggc tgcagtgcgc ggcctccgag
      781 ccgtgccggg cggtcttcag ggaggctgaa gtgaccttgg aggcgggagg cgcggagcag
      841 gagcccggcc aggcgctggg gaaagtattc atgggctgcc ctgggcaaga gccagctctg
      901 tttagcactg ataatgatga cttcactgtg cggaatggcg agacagtcca ggaaagaagg
      961 tcactgaagg aaaggaatcc attgaagatc ttcccatcca aacgtatctt acgaagacac
     1021 aagagagatt gggtggttgc tccaatatct gtccctgaaa atggcaaggg tcccttcccc
     1081 cagagactga atcagctcaa gtctaataaa gatagagaca ccaagatttt ctacagcatc
     1141 acggggccgg gggcagacag cccccctgag ggtgtcttcg ctgtagagaa ggagacaggc
     1201 tggttgttgt tgaataagcc actggaccgg gaggagattg ccaagtatga gctctttggc
     1261 cacgctgtgt cagagaatgg tgcctcagtg gaggacccca tgaacatctc catcatcgtg
     1321 accgaccaga atgaccacaa gcccaagttt acccaggaca ccttccgagg gagtgtctta
     1381 gagggagtcc taccaggtac ttctgtgatg caggtgacag ccacggatga ggatgatgcc
     1441 atctacacct acaatggggt ggttgcttac tccatccata gccaagaacc aaaggaccca
     1501 cacgacctca tgttcaccat tcaccggagc acaggcacca tcagcgtcat ctccagtggc
     1561 ctggaccggg aaaaagtccc tgagtacaca ctgaccatcc aggccacaga catggatggg
     1621 gacggctcca ccaccacggc agtggcagta gtggagatcc ttgatgccaa tgacaatgct
     1681 cccatgtttg acccccagaa gtacgaggcc catgtgcctg agaatgcagt gggccatgag
     1741 gtgcagaggc tgacggtcac tgatctggac gcccccaact caccagcgtg gcgtgccacc
     1801 taccttatca tgggcggtga cgacggggac cattttacca tcaccaccca ccctgagagc
     1861 aaccagggca tcctgacaac caggaagggt ttggattttg aggccaaaaa ccagcacacc
     1921 ctgtacgttg aagtgaccaa cgaggcccct tttgtgctga agctcccaac ctccacagcc
     1981 accatagtgg tccacgtgga ggatgtgaat gaggcacctg tgtttgtccc accctccaaa
     2041 gtcgttgagg tccaggaggg catccccact ggggagcctg tgtgtgtcta cactgcagaa
     2101 gaccctgaca aggagaatca aaagatcagc taccgcatcc tgagagaccc agcagggtgg
     2161 ctagccatgg acccagacag tgggcaggtc acagctgtgg gcaccctcga ccgtgaggat
     2221 gagcagtttg tgaggaacaa catctatgaa gtcatggtct tggccatgga caatggaagc
     2281 cctcccacca ctggcacggg aacccttctg ctaacactga ttgatgtcaa tgaccatggc
     2341 ccagtccctg agccccgtca gatcaccatc tgcaaccaaa gccctgtgcg ccaggtgctg
     2401 aacatcacgg acaaggacct gtctccccac acctcccctt tccaggccca gctcacagat
     2461 gactcagaca tctactggac ggcagaggtc aacgaggaag gtgacacagt ggtcttgtcc
     2521 ctgaagaagt tcctgaagca ggatacatat gacgtgcacc tttctctgtc tgaccatggc
     2581 aacaaagagc agctgacggt gatcagggcc actgtgtgcg actgccatgg ccatgtcgaa
     2641 acctgccctg gaccctggaa gggaggtttc atcctccctg tgctgggggc tgtcctggct
     2701 ctgctgttcc tcctgctggt gctgcttttg ttggtgagaa agaagcggaa gatcaaggag
     2761 cccctcctac tcccagaaga tgacacccgt gacaacgtct tctactatgg cgaagagggg
     2821 ggtggcgaag aggaccagga ctatgacatc acccagctcc accgaggtct ggaggccagg
     2881 ccggaggtgg ttctccgcaa tgacgtggca ccaaccatca tcccgacacc catgtaccgt
     2941 cctcggccag ccaacccaga tgaaatcggc aactttataa ttgagaacct gaaggcggct
     3001 aacacagacc ccacagcccc gccctacgac accctcttgg tgttcgacta tgagggcagc
     3061 ggctccgacg ccgcgtccct gagctccctc acctcctccg cctccgacca agaccaagat
     3121 tacgattatc tgaacgagtg gggcagccgc ttcaagaagc tggcagacat gtacggtggc
     3181 ggggaggacg acctcgagca ccatcaccat caccatcacc attagggccc gatatctcga
     3241 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3301 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3361 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3421 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3481 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3541 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3601 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     3661 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3721 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3781 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     3841 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3901 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     3961 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4021 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4081 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     4141 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4201 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4261 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     4321 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4381 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4441 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4501 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4561 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4621 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4681 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4741 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4801 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4861 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4921 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4981 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5041 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5101 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5161 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5221 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5281 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5341 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5401 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5461 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5521 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5581 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5641 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5701 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5761 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     5821 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5881 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5941 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6001 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6061 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6121 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6181 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6241 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6301 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6361 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6421 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6481 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6541 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6601 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6661 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6721 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6781 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6841 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6901 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6961 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7021 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7081 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7141 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7201 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7261 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7321 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7381 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7441 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7501 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7561 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7621 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7681 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7741 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7801 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7861 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7921 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7981 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8041 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8101 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8161 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8221 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8281 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     8341 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     8401 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     8461 cttagg
//
