LOCUS       V025373                 6472 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025373
VERSION     V025373
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..1965
                     /label="Aurkb(NM_011496)"
                     /note="Aurkb(NM_011496)"
                     /gene="Aurkb"
     CDS             1975..1992
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    2039..2263
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2309..2737
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2751..3080
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3147..3941
                     /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    4115..4236
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4285..4301)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4309..4325
                     /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(4333..4363)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4378..4399
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4687..5272)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5443..6303)
                     /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(6304..6408)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atggctcaga aggagaacgc ctacccgtgg
      961 ccctacggct caaagacgtc tcagtctggc ctgaacacgt tgtcccagag agtcctacgg
     1021 aaggagcccg ccacgacatc tgcgcttgct ctcgtgaacc ggttcaacag ccagtccaca
     1081 gctgcccctg gccagaagtt ggctgagaac aagagtcagg gctccactgc ctcgcaagga
     1141 tcccagaaca agcagccttt cactattgac aactttgaga ttgggcgtcc tttgggcaaa
     1201 ggcaaatttg gaaacgtgta cttggctcgg gagaagaaga gccgtttcat cgtggcactc
     1261 aagatcctct tcaagtctca gattgagaag gagggggtag agcaccagct tcgccgagag
     1321 atcgaaatcc aggcgcacct gaaacatccc aacatccttc aactctacaa ctacttctac
     1381 gaccagcaga ggatctactt aatcctggaa tacgcccctc gcggggaact ctacaaggaa
     1441 ctgcagaaga gtcggacctt cgatgagcag cggactgcca cgatcatgga ggaactgtca
     1501 gatgccctga cctactgcca caagaagaag gtaattcaca gagacataaa gccggagaac
     1561 ctgctgttag gtctgcaggg agaactgaag attgcagact ttggctggtc ggtgcatgcc
     1621 ccatccctga ggaggaagac catgtgcggc acgctggact atctgccccc agagatgatt
     1681 gaggggcgca tgcataatga aatggtagat ctatggtgca tcggggtgct ctgctatgaa
     1741 ctgatggtgg ggaacccacc cttcgagagc cctagccaca gtgagacgta tcgtcggatt
     1801 gtcaaggtgg acctgaagtt cccctcttct gtgccttcgg gcgcccagga cctcatctcc
     1861 aagctgctca aacataaccc ctggcaacgg ctgcccctgg cggaggttgc agctcaccct
     1921 tgggtccggg ccaactcaag gagggttctg cctccctctg cccttgcggc cgcacatcac
     1981 catcatcacc attgactcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     2041 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2101 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2161 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2221 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2281 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2341 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2401 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2461 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2521 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2581 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2641 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2701 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2761 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2821 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2881 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2941 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3001 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3061 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3121 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3181 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3241 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3301 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3361 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3421 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3481 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3541 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3601 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3661 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3721 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3781 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3841 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3901 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3961 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4021 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4081 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4141 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4201 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4261 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4321 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4381 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4441 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4501 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4561 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4621 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4681 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4741 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4801 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4861 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4921 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4981 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5041 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5101 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5161 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5221 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5281 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5341 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5401 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5461 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5521 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5581 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5641 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5701 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5761 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5821 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5881 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5941 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6001 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6061 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6121 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6181 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6241 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6301 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6361 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6421 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gt
//
