LOCUS       V018298                 8179 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018298
VERSION     V018298
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             932..3658
                     /label="HACE1(NM_020771)"
                     /note="HACE1(NM_020771)"
                     /gene="HACE1"
     CDS             3680..3745
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3774..3998
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4044..4472
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4486..4815
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4882..5676
                     /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    5852..5973
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6022..6038)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6046..6062
                     /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(6070..6100)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6115..6136
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6424..7012)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7183..8043)
                     /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(8044..8148)
                     /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 gctggctagt
      901 taagcttggt accgagctcg gatccgccac catggagaga gcgatggagc aactcaaccg
      961 cctgacgcgc tcgctgcgcc gcgcgcgcac cgtggagttg cccgaggata atgaaactgc
     1021 tgtttataca ttaatgccaa tggttatggc tgatcaacac aggtctgttt ctgaactact
     1081 atcaaattca aaatttgatg tcaattatgc attcggacgt gtgaaaagaa gcttgcttca
     1141 cattgcagca aattgtggat cggtggaatg cttggttttg ctgttaaaga aaggagcaaa
     1201 tcctaactat caagatattt caggctgtac accccttcat ttggcagcaa gaaatgggca
     1261 gaagaaatgt atgagtaaat tattagaata tagcgctgat gtcaacattt gtaataatga
     1321 aggccttaca gcaatacatt ggctggctgt gaatgggcgg acagaactac tccatgacct
     1381 tgtgcagcat gtcagtgatg ttgatgttga ggatgccatg gggcagacag cactgcatgt
     1441 tgcctgccag aacggtcaca agacgacagt gcagtgcttg ctagacagtg gtgctgatat
     1501 taacaggcca aatgtatcag gagcaactcc attgtacttt gcttgcagtc atggtcagag
     1561 agatacagca cagatcctac tattacgagg agccaaatat ctgccagata aaaatggagt
     1621 aactcctctg gatttatgtg tacagggtgg atatggagag acttgtgaag tattaattca
     1681 atatcacccg aggctttttc agactattat tcaaatgaca cagaatgaag acctccgaga
     1741 aaacatgtta cggcaagttc tggagcattt gtctcagcaa agtgaaagcc agtacctaaa
     1801 gattctaaca agccttgctg aagttgctac aacaaatggt cataaactgc ttagcctctc
     1861 tagcaattat gatgctcaaa tgaagagcct tttaaggatt gtgagaatgt tttgtcacgt
     1921 ctttcgaatt ggtccatcct cccccagtaa tggaattgat atgggctaca atgggaataa
     1981 aactccaaga agccaggtgt tcaagcctct ggaattgctt tggcactcgt tagatgaatg
     2041 gctagtttta atagccacag aattgatgaa aaacaaaaga gactcaacag agatcacttc
     2101 tattttactg aaacaaaaag gccaagatca agatgctgct tccattcctc catttgaacc
     2161 tccaggacct gggagctatg aaaatctgtc cactggcaca agggaatcta aaccagatgc
     2221 tcttgcaggg agacaggaag ccagtgcaga ttgtcaggat gttatttcta tgacagctaa
     2281 ccggctaagt gctgtcattc aagcttttta catgtgctgt tcttgtcaga tgcctccggg
     2341 aatgacttca cctcgtttca ttgaatttgt ctgcaaacat gatgaagttt taaaatgctt
     2401 tgttaataga aatcccaaaa ttatatttga ccactttcac tttctccttg aatgtcctga
     2461 gttgatgtca agattcatgc atatcataaa agcacagcct tttaaagatc gctgtgaatg
     2521 gttctatgaa catttgcatt caggacagcc agattcagat atggtgcaca ggccagtgaa
     2581 tgaaaatgat atcctgctgg ttcacagaga ttctattttt aggagtagct gtgaagttgt
     2641 gtcaaaagca aattgtgcaa agctaaagca agggattgct gtacggttcc atggagaaga
     2701 aggcatgggt caaggtgttg tgcgtgagtg gtttgatatt ctgtccaatg agatagtcaa
     2761 tcctgattat gcattgttta cccagtcagc tgatggaaca acttttcagc ctaatagcaa
     2821 ctcttatgta aatcctgatc acttgaacta ttttcggttt gctgggcaga tcttgggatt
     2881 agcgttgaac cacaggcagc tggtcaatat ttacttcaca cgatccttct acaagcacat
     2941 tcttggtatt cctgtaaatt accaagatgt ggcatccatt gatccagaat atgcgaaaaa
     3001 tttgcaatgg attttagata atgatataag tgatctgggt ctagaactaa ctttttctgt
     3061 tgagactgat gtgtttggag caatggaaga ggtgcctttg aaacctgggg gtgggagtat
     3121 tcttgtgaca caaaataata aagcggagta cgtccagctt gttactgaac ttcgaatgac
     3181 aagagccatt cagcctcaga tcaatgcttt tttacagggc tttcatatgt tcattccacc
     3241 ctccctcata cagctttttg atgaatatga attggagcta ctgctttctg gcatgccaga
     3301 aattgatgtg agtgattgga taaaaaatac agaatacaca agtggctatg aaagagaaga
     3361 tccagttatt cagtggttct gggaagttgt agaagacatt actcaagagg agagagttct
     3421 tctcttacag tttgttacgg gcagttccag ggtcccacat ggtgggtttg ctaatatcat
     3481 gggtggaagt ggattgcaaa actttacaat cgctgctgtg ccatatactc caaatctttt
     3541 accaacttca agcacatgca tcaacatgct caagttacct gaatacccaa gtaaagaaat
     3601 actcaaggac agacttcttg tggcactaca ttgtggcagc tatggttaca caatggcact
     3661 cgagtctaga gggcccttcg actacaaaga ccatgacggt gattataaag atcatgacat
     3721 cgactacaag gatgacgatg acaagtgagt ttaaacccgc tgatcagcct cgactgtgcc
     3781 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3841 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3901 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3961 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4021 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4081 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4141 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4201 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4261 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4321 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4381 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4441 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4501 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4561 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4621 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4681 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4741 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4801 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4861 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4921 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4981 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5041 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5101 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5161 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5221 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5281 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5341 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5401 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5461 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5521 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5581 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5641 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     5701 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5761 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5821 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5881 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5941 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6001 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6061 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6121 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6181 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6241 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6301 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6361 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6421 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6481 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6541 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6601 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6661 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6721 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6781 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6841 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6901 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6961 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7021 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7081 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7141 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7201 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7261 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7321 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7381 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7441 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7501 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7561 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7621 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7681 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7741 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7801 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7861 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7921 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7981 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8041 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8101 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8161 aaaagtgcca cctgacgtc
//
