LOCUS       V027285                 9120 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027285
VERSION     V027285
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3800
                     /label="NCAPG(NM_022346)"
                     /note="NCAPG(NM_022346)"
                     /gene="NCAPG"
     CDS             3807..3872
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3896..4484
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4550..4598
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4800..5228
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5242..5571
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5638..6432
                     /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    6608..6729
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6778..6794)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6802..6818
                     /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(6826..6856)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6871..6892
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7180..7768)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7939..8799)
                     /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(8800..8904)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggg agcggaaagg aggctgctgt
      781 cgattaagga ggcctttcgg ctggcgcagc agccgcacca gaaccaggcg aagctggtgg
      841 tggcgctgag ccgcacctac cgcacgatgg atgataagac agtttttcat gaggagttca
      901 ttcattacct taaatatgtt atggtggtct ataaacgtga accagctgtg gagagggtaa
      961 tagaatttgc agcaaagttt gttacctcat ttcaccaatc agatatggaa gatgatgagg
     1021 aagaggaaga tggtggcctt ttaaattatt tgtttacttt tctcttaaag tctcatgaag
     1081 caaacagcaa tgcagtgaga tttagagtgt gcctgctcat aaacaagctt ttgggaagta
     1141 tgccagaaaa tgctcagatt gatgatgatg tgtttgataa aattaataaa gccatgctta
     1201 ttagattgaa agataagatt ccaaatgtga gaatacaggc agttctggcg ctttcacgac
     1261 ttcaggatcc caaggatgat gaatgcccag tggttaatgc atatgctact ttgattgaaa
     1321 atgattcaaa tccagaagtt agacgggcag tgttatcatg tattgcacca tcagcaaaga
     1381 ctttgccaaa aattgtaggg cgcaccaagg atgtgaaaga ggctgtcaga aagctggctt
     1441 atcaggtttt agctgaaaag gttcatatga gagctatgtc cattgctcag agagtaatgc
     1501 tccttcaaca aggtcttaat gacagatcag atgctgtgaa acaagctatg cagaagcatc
     1561 ttcttcaagg ctggttacgg ttctctgaag gaaatatctt agagttgctc catcggttgg
     1621 atgtagaaaa ttcttctgaa gtggcagtct ctgttctcaa tgccttgttt tcaataactc
     1681 ctctcagtga actggtggga ctctgtaaaa acaatgatgg caggaaattg attccagtgg
     1741 aaacattaac tcctgaaatt gctttgtatt ggtgtgccct ttgtgaatat ttgaaatcaa
     1801 aaggagatga aggtgaagaa tttttagagc agattttgcc agagcctgta gtatatgcag
     1861 actatttatt gagttacatc cagagcattc cagttgttaa tgaagaacac agaggtgatt
     1921 tttcctatat tggaaatttg atgacaaaag aattcatagg tcaacaattg attctaatta
     1981 ttaagtcttt ggataccagt gaagaaggag gaagaaaaaa actgctggct gttttacagg
     2041 agattcttat tttacccaca atcccaatat ccctggtttc ttttcttgtt gaaagactac
     2101 tccacatcat tatagatgat aataagagaa cacaaattgt tacagaaatt atctcagaga
     2161 ttcgggcgcc cattgttact gttggtgtta ataacgatcc agctgatgta agaaagaaag
     2221 aactcaagat ggctgaaata aaagttaagc ttatcgaagc caaagaagct ttggaaaatt
     2281 gcattacctt acaggatttt aatcgggcat cagaattaaa agaagaaata aaagcattag
     2341 aagatgccag aataaacctt ttgaaagaga cagagcaact tgaaattaaa gaagtccaca
     2401 tagagaagaa tgatgctgaa acattgcaga aatgtcttat tttatgctat gaactgttga
     2461 agcagatgtc catttcaaca ggcttaagtg caaccatgaa tggaatcatc gaatctttga
     2521 ttcttcctgg aataataagt attcatcctg ttgtaagaaa cctggctgtt ttatgcttgg
     2581 gatgctgtgg actacagaat caggattttg caaggaaaca cttcgtatta ctattgcagg
     2641 ttttgcaaat tgatgatgtc acaataaaaa taagtgcttt aaaggcaatc tttgaccaac
     2701 tgatgacgtt cgggattgaa ccatttaaaa ctaaaaaaat caaaacactt cattgtgaag
     2761 gtacagaaat aaacagtgat gatgagcaag aatcaaaaga agttgaagag actgctacag
     2821 ctaagaatgt tctgaaactc ctttctgatt tcttagatag tgaggtatct gaacttagga
     2881 ctggagctgc agaaggacta gccaagctga tgttctctgg gcttttggtc agcagcagga
     2941 ttctttctcg tcttattttg ttatggtaca atcctgtgac tgaagaggat gttcaacttc
     3001 gacattgcct aggcgtgttc ttccccgtgt ttgcttatgc aagcaggact aatcaggaat
     3061 gctttgaaga agcttttctt ccaaccctgc aaacactggc caatgcccct gcatcttctc
     3121 ctttagctga aattgatatc acaaatgttg ctgagttact tgtagatttg acaagaccaa
     3181 gtggattaaa tcctcaggcc aagacttccc aagattatca ggccttaaca gtacatgaca
     3241 atttggctat gaaaatttgc aatgagatct taacaagtcc gtgctcgcca gaaattcgag
     3301 tctatacaaa agccttgagt tctttagaac tcagtagcca tcttgcaaaa gatcttctgg
     3361 ttctattgaa tgagattctg gagcaagtaa aagataggac atgtctgaga gctttggaga
     3421 aaatcaagat tcagttagaa aaaggaaata aagaatttgg tgaccaagct gaagcagcac
     3481 aggatgccac cttgactaca actactttcc aaaatgaaga tgaaaagaat aaagaagtat
     3541 atatgactcc actcaggggt gtaaaagcaa cccaagcatc aaagtctact cagctaaaga
     3601 ctaacagagg acagagaaaa gtgacagttt cagctaggac gaacaggagg tgtcagactg
     3661 ctgaagccga ctctgaaagt gatcatgaag ttccagaacc agaatcagaa atgaagatga
     3721 gactaccaag acgagccaaa accgcagcac tagaaaaaag taaacttaac cttgcccaat
     3781 ttctcaatga agatctaagt ggatccgact acaaagacca tgacggtgat tataaagatc
     3841 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     3901 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3961 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     4021 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     4081 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     4141 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     4201 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     4261 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     4321 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4381 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4441 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     4501 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     4561 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4621 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4681 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4741 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     4801 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4861 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4921 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     4981 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5041 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5101 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5161 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5221 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5281 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5341 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5401 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5461 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5521 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     5581 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     5641 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5701 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5761 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5821 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5881 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5941 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     6001 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     6061 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     6121 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     6181 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     6241 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     6301 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     6361 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6421 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     6481 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6541 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6601 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     6661 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6721 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     6781 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6841 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6901 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6961 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     7021 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7081 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7141 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7201 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7261 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7321 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     7381 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7441 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7501 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7561 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7621 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7681 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7741 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7801 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7861 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7921 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7981 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     8041 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8101 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8161 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8221 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8281 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8341 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8401 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8461 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8521 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8581 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8641 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8701 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8761 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8821 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8881 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     8941 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     9001 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     9061 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
