LOCUS       V023560                 7814 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023560
VERSION     V023560
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             941..3304
                     /label="CDH10(NM_006727)"
                     /note="CDH10(NM_006727)"
                     /gene="CDH10"
     CDS             3311..3376
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3414..3638
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3684..4112
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4126..4455
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4522..5316
                     /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    5490..5611
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5660..5676)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5684..5700
                     /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(5708..5738)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5753..5774
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6062..6647)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6818..7678)
                     /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(7679..7783)
                     /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 atccgccacc atgacaatac atcaattttt
      961 gctactgttt ctattctggg tatgcctgcc acatttctgc tctccagaaa taatgttcag
     1021 aaggacgcct gtgccacagc aaagaatttt aagttcacgt gtaccaagga gtgatggcaa
     1081 aattctccat cgtcaaaaac gtggttggat gtggaatcaa tttttcttac ttgaagaata
     1141 tacaggatct gattatcagt acgtaggcaa gctacattca gaccaagata aaggagatgg
     1201 atcactcaaa tatatcttat ctggagatgg agctggtact ctttttatta ttgatgaaaa
     1261 aacaggtgat attcatgcca caaggcgaat tgatagggag gaaaaggcct tttatactct
     1321 acgcgcacaa gctattaaca gaagaactct gaggccagta gagccagagt cagagtttgt
     1381 gatcaaaatt catgatatca atgacaatga gccaacgttc ccagaagaaa tctatacagc
     1441 tagtgttccc gaaatgtctg ttgtaggtac ttctgtggtg caagtcacag ctacagatgc
     1501 cgatgaccct tcatatggga acagcgccag agtcatttac agcatacttc aagggcagcc
     1561 ctatttctct gtggagcctg aaacaggtat catcaggact gctttaccga acatgaacag
     1621 agaaaacaga gagcaatacc aagtggtcat ccaggccaaa gacatgggcg gccagatggg
     1681 aggcttatcg gggacaacca ctgtgaacat cacgctgaca gatgtcaatg acaacccacc
     1741 acgtttcccc cagaacacta ttcatcttcg agttcttgaa tcctccccag ttggcacagc
     1801 cattggaagt gtcaaagcaa ctgatgctga cactgggaaa aatgctgaag tagaataccg
     1861 aattattgat ggtgacggta ctgatatgtt tgacatcgtg actgagaagg acacacagga
     1921 aggcatcatc actgtgaaaa agccactcga ctatgagagc cgaagacttt atactctgaa
     1981 agtcgaagca gaaaacaccc atgtagatcc ccgtttttat tacctaggac catttaaaga
     2041 tactaccata gtgaaaatct ctatagaaga tgtggatgaa cctcctgttt ttagtaggtc
     2101 ctcctatctg tttgaagttc atgaagatat tgaagtgggc acaatcattg gtactgtaat
     2161 ggcaagggac ccagattcta tttccagccc cattagattt tccttggatc gccatactga
     2221 ccttgacaga atctttaaca ttcattcagg aaatggatct ctttatacat caaaacctct
     2281 tgaccgtgaa ctatctcagt ggcataatct tactgttatt gctgctgaaa tcaacaatcc
     2341 caaagagaca acacgcgtgg ctgtttttgt gagaattttg gatgttaatg acaatgcccc
     2401 acagtttgct gtgttctatg acacttttgt atgtgaaaat gccagaccag ggcagctaat
     2461 acagactata agtgcagtag acaaagatga ccctttaggt ggacagaaat tttttttcag
     2521 tttagctgct gtcaatccaa acttcacagt acaggataat gaagataata ctgccagaat
     2581 cttaaccaga aaaaatggat tcaatagaca tgaaatcagt acctatctct tgcctgtggt
     2641 gatatcagac aatgattacc caattcagag cagcacaggc acactgacca ttcgagtgtg
     2701 tgcttgtgac agccaaggca acatgcaatc ctgcagtgct gaagccctgc tcctccctgc
     2761 cggcctcagc actggggcct tgatcgccat cctcctctgc atcatcattc tactggttat
     2821 agtagtactg tttgcagctc tgaaaagaca gcgaaaaaaa gagcctctga tcttgtcaaa
     2881 agaagatatc agagacaaca ttgtgagcta taacgatgag ggtggtggag aggaggacac
     2941 ccaggccttt gatatcggca ccctgaggaa tcctgcagcc attgaggaaa aaaagctccg
     3001 gcgagatatt attccagaaa cgttatttat tcctcggagg actcctacag ctccagataa
     3061 cacggacgtc cgggatttca ttaatgaaag gctaaaagag catgatcttg accccaccgc
     3121 acccccctac gactcacttg caacctatgc ctatgaagga aatgattcca ttgctgaatc
     3181 tctgagttca ttagaatcag gtactactga aggagaccaa aactacgatt acctccgaga
     3241 atggggccct cggtttaata agctagcaga aatgtatggt ggtggggaaa gtgacaaaga
     3301 ctctctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     3361 ggatgacgat gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     3421 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3481 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3541 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3601 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3661 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3721 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3781 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3841 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3901 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3961 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4021 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4081 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4141 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4201 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4261 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4321 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4381 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4441 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4501 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4561 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4621 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4681 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4741 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4801 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4861 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4921 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4981 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5041 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5101 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5161 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5221 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5281 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5341 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5401 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5461 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5521 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5581 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5641 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5701 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5761 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5821 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5881 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5941 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6001 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6061 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt
     6601 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6661 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6721 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6781 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6841 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6901 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6961 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7021 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7081 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7141 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7201 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7261 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7321 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7381 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7441 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7501 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7561 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7621 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7681 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7741 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7801 tgccacctga cgtc
//
