LOCUS       V031321                 7150 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031321
VERSION     V031321
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             964..2640
                     /label="BORA(NM_024808)"
                     /note="BORA(NM_024808)"
                     /gene="BORA"
     CDS             2647..2712
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2750..2974
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3020..3448
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3462..3791
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3858..4652
                     /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    4826..4947
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4996..5012)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5020..5036
                     /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(5044..5074)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5089..5110
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5398..5983)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6154..7014)
                     /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(7015..7119)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatgggag atgtcaagga atcaaagatg caaataacac cagaaactcc aggaaggatc
     1021 cctgttttaa atccttttga aagtcctagt gattattcta atctccatga acaaactctc
     1081 gccagtcctt ctgtttttaa atcaacaaaa ttaccaactc cagggaaatt tagatggtct
     1141 attgatcaac tagctgtaat aaatcctgta gaaatagacc cagaagatat tcatcgtcaa
     1201 gctttatact taagtcattc tcgaatagat aaagatgtgg aagacaaaag acaaaaagcc
     1261 attgaagagt ttttcactaa agatgtcatc gtaccctctc cttggactga tcatgaaggg
     1321 aaacagcttt cacaatgtca ttccagtaaa tgcactaaca taaatagtga ctctccagtt
     1381 ggaaaaaagc tgaccattca ttctgagaaa agcgatgctg cttgtcagac attgctgtct
     1441 cttcctgtgg attttaattt agaaaatata ttaggtgact attttagagc tgatgaattt
     1501 gcagatcaat ctcctggaaa cctcagttct tcatccctca gaagaaagct gtttttagat
     1561 gggaacggaa gcatctccga ctccttacct tcggcttctc ccggaagtcc tcacagtggt
     1621 gttcaaacat cactagagat gttttattca atagatttgt ctcctgtaaa gtgtaggagc
     1681 cccttgcaga caccaagttc ggggcagttt tcttctagcc ctattcaggc tagtgcaaaa
     1741 aaatacagct tgggaagcat aactagtcct tcgcctattt cttcacccac tttctcacca
     1801 attgaatttc agataggaga gactccactc tcagaacaaa ggaagtttac tgttcattct
     1861 cctgatgctt catctggaac aaattctaat gggataacta atccgtgtat cagaagtcct
     1921 tatatagatg gctgctcgcc aattaaaaat tggtctccta tgagacttca gatgtatagt
     1981 ggtggtactc agtatcggac ctcagtgatt cagatacctt ttactcttga gactcaaggt
     2041 gaagatgagg aagataaaga gaatattcct tccacagatg tctcatcacc cgccatggat
     2101 gctgctggaa tacacctacg gcagtttagt aatgaggctt ctacccatgg tacacatttg
     2161 gttgtgactg ccatgtctgt tacacaaaat cagtccagtg cttctgagaa agaattagca
     2221 ctgttgcagg atgttgaaag ggagaaagac aataacactg tggatatggt tgatcctata
     2281 gagatagcag atgagaccac ttggattaag gagccggttg ataatggcag tttacccatg
     2341 actgattttg taagtggcat tgccttcagt attgaaaact ctcatatgtg catgtcacct
     2401 cttgctgaaa gcagtgtcat tccttgtgaa agcagtaaca ttcagatgga tagtggctat
     2461 aatacgcaga attgtggaag caatattatg gatacagttg gggcagaaag ttactgcaaa
     2521 gaaagtgatg cacaaacatg tgaagttgag agtaaatctc aagcatttaa tatgaagcaa
     2581 gaccacacaa cacagaggtg ttggatgaaa acagcaagcc cttttcaatg cagcagtcca
     2641 ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat
     2701 gacgatgaca agtagtgagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     2761 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2821 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2881 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2941 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3001 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3061 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3121 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3181 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3241 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3301 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3361 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3421 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3481 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3541 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3601 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3661 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3721 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3781 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     3841 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3901 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3961 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4021 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4081 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4141 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4201 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4261 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4321 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4381 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4441 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4501 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4561 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4621 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4681 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4741 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4801 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4861 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4921 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4981 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5041 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5101 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5161 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5221 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5281 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5341 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5401 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5461 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5521 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5581 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5641 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5701 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5761 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5821 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5881 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     5941 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6001 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6061 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6121 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6181 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6241 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6301 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6361 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6421 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6481 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6541 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6601 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6661 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6721 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6781 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6841 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6901 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6961 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7021 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7081 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7141 acctgacgtc
//
