LOCUS       V025778                 6167 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025778
VERSION     V025778
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..1657
                     /label="DIABLO(NM_001371333)"
                     /note="DIABLO(NM_001371333)"
                     /gene="DIABLO"
     CDS             1664..1729
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    1767..1991
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2037..2465
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2479..2808
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2875..3669
                     /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    3843..3964
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4013..4029)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4037..4053
                     /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(4061..4091)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4106..4127
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4415..5000)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5171..6031)
                     /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(6032..6136)
                     /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 atggcggctc tgaagagttg
      961 gctgtcgcgc agcgtaactt cattcttcag gtacagacag tgtttgtgtg ttcctgttgt
     1021 ggctaacttt aagaagcggt gtttctcaga attgataaga ccatggcaca aaactgtgac
     1081 gattggcttt ggagtaaccc tgtgtgcggt tcctattgca cagaaatcag agcctcattc
     1141 ccttagtagt gaagcattga tgaggagagc agtgtctttg gtaacagata gcacctctac
     1201 ctttctctct cagaccacat atgcgttgat tgaagctatt actgaatata ctaaggctgt
     1261 ttatacctta acttctcttt accgacaata tacaagttta cttgggaaaa tgaattcaga
     1321 ggaggaagat gaagtgtggc aggtgatcat aggagccaga gctgagatga cttcaaaaca
     1381 ccaagagtac ttgaagctgg aaaccacttg gatgactgca gttggtcttt cagagatggc
     1441 agcagaagct gcatatcaaa ctggcgcaga tcaggcctct ataaccgcca ggaatcacat
     1501 tcagctggtg aaactgcagg tggaagaggt gcaccagctc tcccggaaag cagaaaccaa
     1561 gctggcagaa gcacagatag aagagctccg tcagaaaaca caggaggaag gggaggagcg
     1621 ggctgagtcg gagcaggagg cctacctgcg tgaggatctc gaggactaca aagaccatga
     1681 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     1741 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     1801 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1861 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1921 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     1981 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     2041 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2101 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2161 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2221 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2281 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2341 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2401 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2461 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2521 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2581 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2641 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2701 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     2761 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     2821 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2881 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2941 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3001 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3061 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3121 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3181 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3241 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3301 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3361 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3421 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3481 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3541 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3601 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3661 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3721 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3781 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3841 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3901 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3961 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4021 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4081 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4141 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4201 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4261 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4321 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4381 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4441 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4501 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4561 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4621 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4681 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4741 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4801 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4861 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4921 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     4981 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5041 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5101 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5161 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5221 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5281 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5341 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5401 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5461 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5521 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5581 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5641 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5701 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5761 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5821 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5881 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5941 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6001 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6061 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6121 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
