LOCUS       V030348                 8745 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030348
VERSION     V030348
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             681..698
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     CDS             708..3455
                     /label="CDH4(NM_001794)"
                     /note="CDH4(NM_001794)"
                     /gene="CDH4"
     misc_feature    3521..4109
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4175..4223
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4425..4853
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4867..5196
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5263..6057
                     /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    6233..6354
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6403..6419)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6427..6443
                     /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(6451..6481)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6496..6517
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6805..7393)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7564..8424)
                     /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(8425..8529)
                     /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 ttaagcagat
      661 atccagcaca ggccaccatg caccatcacc atcaccatct tggtaccatg accgcgggcg
      721 ccggcgtgct ccttctgctg ctctcgctct ccggcgcgct ccgggcccat aatgaggatc
      781 ttacaactag agagacctgc aaggctgggt tctctgaaga tgattacacg gcattaatct
      841 cccaaaatat tctagaaggg gaaaagctac ttcaagtcaa gttcagcagc tgtgtgggga
      901 ccaaggggac acaatatgag accaacagca tggacttcaa agttggggca gatgggacag
      961 tcttcgccac ccgggagctg caggtcccct ccgagcaggt ggcgttcacg gtgactgcat
     1021 gggacagcca gacagcagag aaatgggacg ccgtggtgcg gttgctggtg gcccagacct
     1081 cgtccccgca ctctggacac aagccgcaga aaggaaagaa ggtcgtggct ctggacccct
     1141 ctccgcctcc gaaggacacc ctgctgccgt ggccccagca ccagaacgcc aacgggctga
     1201 ggcggcgcaa acgggactgg gtcatcccgc ccatcaacgt gcccgagaac tcgcgcgggc
     1261 ccttcccgca gcagctcgtg aggatccggt ccgacaaaga caatgacatc cccatccggt
     1321 acagcatcac gggagtgggc gccgaccagc cccccatgga ggtcttcagc attgactcca
     1381 tgtccggccg gatgtacgtc acaaggccca tggaccggga ggagcacgcc tcttaccacc
     1441 tccgagccca cgctgtggac atgaatggca acaaggtgga gaaccccatc gacctgtaca
     1501 tctacgtcat cgacatgaat gacaaccgcc ctgagttcat caaccaggtc tacaacggct
     1561 ccgtggacga gggctccaag ccaggcacct acgtgatgac cgtcacggcc aacgatgctg
     1621 acgacagcac cacggccaac gggatggtgc ggtaccggat cgtgacccag accccacaga
     1681 gcccgtccca gaatatgttc accatcaaca gcgagactgg agatatcgtc acagtggcgg
     1741 ctggcctgga ccgagagaaa gttcagcagt acacagtcat cgttcaggcc acagatatgg
     1801 aaggaaatct caactatggc ctctcaaaca cagccacagc catcatcacg gtgacagatg
     1861 tgaatgacaa cccgccagaa tttaccgcca gcacgtttgc aggggaggtc cccgaaaacc
     1921 gcgtggagac cgtggtcgca aacctcacgg tgatggaccg agatcagccc cactctccaa
     1981 actggaatgc cgtttaccgc atcatcagtg gggatccatc cgggcacttc agcgtccgca
     2041 cagaccccgt aaccaacgag ggcatggtca ccgtggtgaa ggcagtcgac tacgagctca
     2101 acagagcttt catgctgaca gtgatggtgt ccaaccaggc gcccctggcc agcggaatcc
     2161 agatgtcctt ccagtccacg gcaggggtga ccatctccat catggacatc aacgaggctc
     2221 cctacttccc ctcaaaccac aagctgatcc gcctggagga gggcgtgccc cccggcaccg
     2281 tgctgaccac gttttcagct gtggaccctg accggttcat gcagcaggct gtgagatact
     2341 caaagctgtc agacccagcg agctggctgc acatcaatgc caccaacggc cagatcacca
     2401 cggcggcagt gctggaccgt gagtccctct acaccaaaaa caacgtctac gaggccacct
     2461 tcctggcagc tgacaatggg atacccccgg ccagcggcac cgggaccctc cagatctatc
     2521 tcattgacat caacgacaac gcccctgagc tgctgcccaa ggaggcgcag atctgcgaga
     2581 agcccaacct gaacgccatc aacatcacgg cggccgacgc tgacgtcgac cccaacatcg
     2641 gcccctacgt cttcgagctg ccctttgtcc cggcggccgt gcggaagaac tggaccatca
     2701 cccgcctgaa cggtgactat gcccaactca gcttgcgcat cctgtacctg gaggccggga
     2761 tgtatgacgt ccccatcatc gtcacagact ctggaaaccc tcccctgtcc aacacgtcca
     2821 tcatcaaagt caaggtgtgc ccatgtgatg acaacgggga ctgcaccacc attggcgcag
     2881 tggcagcggc tggtctgggc accggtgcca tcgtggccat cctcatctgc atcctcatcc
     2941 tgctgaccat ggtcctgctg tttgtcatgt ggatgaagcg gcgagagaag gagcgccaca
     3001 cgaagcagct gctcattgac cccgaggacg acgtccgcga caacatcctc aagtatgacg
     3061 aggaaggcgg tggcgaggag gaccaggact acgacctcag ccagctgcag cagccggaag
     3121 ccatggggca cgtgccaagc aaagcccctg gcgtgcgtcg cgtggatgag cggccggtgg
     3181 gcgctgagcc ccagtacccg atcaggccca tggtgccgca cccaggcgac atcggtgact
     3241 tcatcaatga gggactccgc gctgctgaca acgaccccac ggcacccccc tatgactccc
     3301 tgctggtctt cgactacgag gggagcggct ccaccgcagg ctccgtcagc tccctgaact
     3361 catccagttc cggggaccaa gactacgatt acctcaacga ctgggggccc agattcaaga
     3421 agctggcgga catgtatgga ggtggtgaag aggattgaga attctgcaga tatccagcac
     3481 agtggcggcc gctcgagtct agagggcccg atatctcgac aatcaacctc tggattacaa
     3541 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3601 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3661 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3721 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3781 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3841 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3901 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3961 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     4021 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4081 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     4141 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4201 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4261 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4321 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4381 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4441 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4501 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4561 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     4621 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4681 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4741 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     4801 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4861 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4921 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4981 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5041 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5101 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     5161 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5221 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5281 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5341 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5401 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5461 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5521 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5581 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5641 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5701 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5761 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5821 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5881 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5941 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6001 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6061 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6121 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6181 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     6241 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6301 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6361 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6421 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6481 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6541 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6601 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6661 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6721 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6781 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6841 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6901 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6961 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     7021 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7081 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7141 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7201 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7261 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7321 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7381 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7441 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7501 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7561 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7621 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7681 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7741 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7801 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7861 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7921 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7981 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8041 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8101 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8161 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8221 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8281 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8341 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8401 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8461 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8521 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     8581 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     8641 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     8701 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
