LOCUS       V037774                10610 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037774
VERSION     V037774
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"
     CDS             683..5275
                     /label="TOP2A(NM_001067)"
                     /note="TOP2A(NM_001067)"
                     /gene="TOP2A"
     CDS             5282..5371
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    5386..5974
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    6040..6088
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6290..6718
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6732..7061
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7128..7922
                     /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    8098..8219
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8268..8284)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8292..8308
                     /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(8316..8346)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8361..8382
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8670..9258)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9429..10289)
                     /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(10290..10394)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggaagt gtcaccattg cagcctgtaa atgaaaatat
      721 gcaagtcaac aaaataaaga aaaatgaaga tgctaagaaa agactgtctg ttgaaagaat
      781 ctatcaaaag aaaacacaat tggaacatat tttgctccgc ccagacacct acattggttc
      841 tgtggaatta gtgacccagc aaatgtgggt ttacgatgaa gatgttggca ttaactatag
      901 ggaagtcact tttgttcctg gtttgtacaa aatctttgat gagattctag ttaatgctgc
      961 ggacaacaaa caaagggacc caaaaatgtc ttgtattaga gtcacaattg atccggaaaa
     1021 caatttaatt agtatatgga ataatggaaa aggtattcct gttgttgaac acaaagttga
     1081 aaagatgtat gtcccagctc tcatatttgg acagctccta acttctagta actatgatga
     1141 tgatgaaaag aaagtgacag gtggtcgaaa tggctatgga gccaaattgt gtaacatatt
     1201 cagtaccaaa tttactgtgg aaacagccag tagagaatac aagaaaatgt tcaaacagac
     1261 atggatggat aatatgggaa gagctggtga gatggaactc aagcccttca atggagaaga
     1321 ttatacatgt atcacctttc agcctgattt gtctaagttt aaaatgcaaa gcctggacaa
     1381 agatattgtt gcactaatgg tcagaagagc atatgatatt gctggatcca ccaaagatgt
     1441 caaagtcttt cttaatggaa ataaactgcc agtaaaagga tttcgtagtt atgtggacat
     1501 gtatttgaag gacaagttgg atgaaactgg taactccttg aaagtaatac atgaacaagt
     1561 aaaccacagg tgggaagtgt gtttaactat gagtgaaaaa ggctttcagc aaattagctt
     1621 tgtcaacagc attgctacat ccaagggtgg cagacatgtt gattatgtag ctgatcagat
     1681 tgtgactaaa cttgttgatg ttgtgaagaa gaagaacaag ggtggtgttg cagtaaaagc
     1741 acatcaggtg aaaaatcaca tgtggatttt tgtaaatgcc ttaattgaaa acccaacctt
     1801 tgactctcag acaaaagaaa acatgacttt acaacccaag agctttggat caacatgcca
     1861 attgagtgaa aaatttatca aagctgccat tggctgtggt attgtagaaa gcatactaaa
     1921 ctgggtgaag tttaaggccc aagtccagtt aaacaagaag tgttcagctg taaaacataa
     1981 tagaatcaag ggaattccca aactcgatga tgccaatgat gcagggggcc gaaactccac
     2041 tgagtgtacg cttatcctga ctgagggaga ttcagccaaa actttggctg tttcaggcct
     2101 tggtgtggtt gggagagaca aatatggggt tttccctctt agaggaaaaa tactcaatgt
     2161 tcgagaagct tctcataagc agatcatgga aaatgctgag attaacaata tcatcaagat
     2221 tgtgggtctt cagtacaaga aaaactatga agatgaagat tcattgaaga cgcttcgtta
     2281 tgggaagata atgattatga cagatcagga ccaagatggt tcccacatca aaggcttgct
     2341 gattaatttt atccatcaca actggccctc tcttctgcga catcgttttc tggaggaatt
     2401 tatcactccc attgtaaagg tatctaaaaa caagcaagaa atggcatttt acagccttcc
     2461 tgaatttgaa gagtggaaga gttctactcc aaatcataaa aaatggaaag tcaaatatta
     2521 caaaggtttg ggcaccagca catcaaagga agctaaagaa tactttgcag atatgaaaag
     2581 acatcgtatc cagttcaaat attctggtcc tgaagatgat gctgctatca gcctggcctt
     2641 tagcaaaaaa cagatagatg atcgaaagga atggttaact aatttcatgg aggatagaag
     2701 acaacgaaag ttacttgggc ttcctgagga ttacttgtat ggacaaacta ccacatatct
     2761 gacatataat gacttcatca acaaggaact tatcttgttc tcaaattctg ataacgagag
     2821 atctatccct tctatggtgg atggtttgaa accaggtcag agaaaggttt tgtttacttg
     2881 cttcaaacgg aatgacaagc gagaagtaaa ggttgcccaa ttagctggat cagtggctga
     2941 aatgtcttct tatcatcatg gtgagatgtc actaatgatg accattatca atttggctca
     3001 gaattttgtg ggtagcaata atctaaacct cttgcagccc attggtcagt ttggtaccag
     3061 gctacatggt ggcaaggatt ctgctagtcc acgatacatc tttacaatgc tcagctcttt
     3121 ggctcgattg ttatttccac caaaagatga tcacacgttg aagtttttat atgatgacaa
     3181 ccagcgtgtt gagcctgaat ggtacattcc tattattccc atggtgctga taaatggtgc
     3241 tgaaggaatc ggtactgggt ggtcctgcaa aatccccaac tttgatgtgc gtgaaattgt
     3301 aaataacatc aggcgtttga tggatggaga agaacctttg ccaatgcttc caagttacaa
     3361 gaacttcaag ggtactattg aagaactggc tccaaatcaa tatgtgatta gtggtgaagt
     3421 agctattctt aattctacaa ccattgaaat ctcagagctt cccgtcagaa catggaccca
     3481 gacatacaaa gaacaagttc tagaacccat gttgaatggc accgagaaga cacctcctct
     3541 cataacagac tatagggaat accatacaga taccactgtg aaatttgttg tgaagatgac
     3601 tgaagaaaaa ctggcagagg cagagagagt tggactacac aaagtcttca aactccaaac
     3661 tagtctcaca tgcaactcta tggtgctttt tgaccacgta ggctgtttaa agaaatatga
     3721 cacggtgttg gatattctaa gagacttttt tgaactcaga cttaaatatt atggattaag
     3781 aaaagaatgg ctcctaggaa tgcttggtgc tgaatctgct aaactgaata atcaggctcg
     3841 ctttatctta gagaaaatag atggcaaaat aatcattgaa aataagccta agaaagaatt
     3901 aattaaagtt ctgattcaga ggggatatga ttcggatcct gtgaaggcct ggaaagaagc
     3961 ccagcaaaag gttccagatg aagaagaaaa tgaagagagt gacaacgaaa aggaaactga
     4021 aaagagtgac tccgtaacag attctggacc aaccttcaac tatcttcttg atatgcccct
     4081 ttggtattta accaaggaaa agaaagatga actctgcagg ctaagaaatg aaaaagaaca
     4141 agagctggac acattaaaaa gaaagagtcc atcagatttg tggaaagaag acttggctac
     4201 atttattgaa gaattggagg ctgttgaagc caaggaaaaa caagatgaac aagtcggact
     4261 tcctgggaaa ggggggaagg ccaaggggaa aaaaacacaa atggctgaag ttttgccttc
     4321 tccgcgtggt caaagagtca ttccacgaat aaccatagaa atgaaagcag aggcagaaaa
     4381 gaaaaataaa aagaaaatta agaatgaaaa tactgaagga agccctcaag aagatggtgt
     4441 ggaactagaa ggcctaaaac aaagattaga aaagaaacag aaaagagaac caggtacaaa
     4501 gacaaagaaa caaactacat tggcatttaa gccaatcaaa aaaggaaaga agagaaatcc
     4561 ctggtctgat tcagaatcag ataggagcag tgacgaaagt aattttgatg tccctccacg
     4621 agaaacagag ccacggagag cagcaacaaa aacaaaattc acaatggatt tggattcaga
     4681 tgaagatttc tcagattttg atgaaaaaac tgatgatgaa gattttgtcc catcagatgc
     4741 tagtccacct aagaccaaaa cttccccaaa acttagtaac aaagaactga aaccacagaa
     4801 aagtgtcgtg tcagaccttg aagctgatga tgttaagggc agtgtaccac tgtcttcaag
     4861 ccctcctgct acacatttcc cagatgaaac tgaaattaca aacccagttc ctaaaaagaa
     4921 tgtgacagtg aagaagacag cagcaaaaag tcagtcttcc acctccacta ccggtgccaa
     4981 aaaaagggct gccccaaaag gaactaaaag ggatccagct ttgaattctg gtgtctctca
     5041 aaagcctgat cctgccaaaa ccaagaatcg ccgcaaaagg aagccatcca cttctgatga
     5101 ttctgactct aattttgaga aaattgtttc gaaagcagtc acaagcaaga aatccaaggg
     5161 ggagagtgat gacttccata tggactttga ctcagctgtg gctcctcggg caaaatctgt
     5221 acgggcaaag aaacctataa agtacctgga agagtcagat gaagatgatc tgtttctcga
     5281 ggaacaaaaa ctaatatcag aggaagattt ggaacagaaa ctaatcagtg aagaagattt
     5341 agagcaaaag ttaatttccg aggaggactt ataggatatc tcgacaatca acctctggat
     5401 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     5461 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     5521 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     5581 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     5641 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     5701 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     5761 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     5821 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     5881 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     5941 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa
     6001 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     6061 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     6121 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     6181 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     6241 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt
     6301 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     6361 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     6421 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg
     6481 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     6541 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     6601 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     6661 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     6721 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6781 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     6841 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     6901 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     6961 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     7021 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     7081 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     7141 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     7201 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     7261 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     7321 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     7381 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     7441 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     7501 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     7561 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     7621 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     7681 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     7741 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     7801 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     7861 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     7921 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     7981 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     8041 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     8101 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     8161 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     8221 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     8281 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     8341 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     8401 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     8461 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     8521 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     8581 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     8641 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     8701 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     8761 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     8821 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     8881 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     8941 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     9001 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     9061 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     9121 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     9181 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     9241 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     9301 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     9361 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     9421 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     9481 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     9541 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     9601 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     9661 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     9721 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     9781 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     9841 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     9901 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     9961 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
    10021 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
    10081 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
    10141 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
    10201 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
    10261 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
    10321 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
    10381 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
    10441 ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc
    10501 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
    10561 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
