LOCUS       V027761                 7758 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027761
VERSION     V027761
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..2438
                     /label="CENPT(NM_025082)"
                     /note="CENPT(NM_025082)"
                     /gene="CENPT"
     CDS             2445..2510
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2534..3122
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3188..3236
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3438..3866
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3880..4209
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4276..5070
                     /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    5246..5367
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5416..5432)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5440..5456
                     /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(5464..5494)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5509..5530
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5818..6406)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6577..7437)
                     /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(7438..7542)
                     /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 ccaccatggc tgaccacaac cctgacagcg
      781 actccacgcc gcgcacgctg ctgcgacgcg tgctggatac agcggacccg cgcaccccgc
      841 ggcgaccccg gagtgctcgg gctggagccc ggagagccct gcttgaaacg gcttccccca
      901 ggaagttgag tggccaaaca aggacgatag ccagagggcg ttcccatgga gccaggtctg
      961 ttggcagatc ggcccatatt caggccagtg ggcacttgga ggaacagaca cctcggacgc
     1021 tgctgaagaa catcctacta actgccccag aatcttccat cctgatgcct gagtcggtag
     1081 tgaagccagt gccagcaccg caggcggtcc aaccctccag acaagagagc agttgcggca
     1141 gcctggagct gcaacttcct gagctcgagc cccccacaac cctggctcca ggtctgctgg
     1201 cccctggcag gaggaaacag aggctgagac tgtcagtgtt tcagcaggga gtggaccagg
     1261 ggctgtctct ctcccaagag cctcaaggga atgctgatgc ctcttccctc accagatccc
     1321 tcaacctgac ctttgccaca cctcttcagc cacagtcagt gcagaggcct ggcttggccc
     1381 gcagacctcc agcccgccga gctgtagacg tgggtgcctt tttgcgggat ctgcgagata
     1441 cttccctggc tcctccaaac attgtgttgg aggacaccca gccgttctct cagcccatgg
     1501 ttggctcccc caacgtgtat cactccctgc cctgcacgcc tcacactggg gctgaagacg
     1561 ctgagcaggc tgccggtcgc aagacacaga gcagtgggcc tgggctgcag aagaatagcc
     1621 ctgggaaacc agcccagttt ctggcaggag aggcagagga ggtcaatgcc tttgctctgg
     1681 gcttcctgag caccagcagt ggtgtctctg gagaagatga agtagagccc ttacacgatg
     1741 gagttgaaga ggcagagaaa aagatggaag aagaaggtgt gagtgtgagt gaaatggagg
     1801 caacaggagc acaaggaccc agcagggtag aagaggctga gggacacaca gaggtgacag
     1861 aagcagaggg atcccagggg actgctgagg ctgacgggcc aggagcatct tcaggggatg
     1921 aggatgcctc tggcagggca gcaagtccag agtcggcctc cagcacccct gagtctctcc
     1981 aggccaggcg acatcatcag tttcttgagc cagccccagc gcctggtgct gcagtcttat
     2041 cttcagagcc tgcagagcct ctgttggtca ggcatccccc taggccccgg accaccggcc
     2101 ccaggccccg gcaagatccc cacaaggctg gactgagcca ctatgtgaaa ctctttagct
     2161 tctatgccaa gatgcccatg gagaggaagg ctcttgagat ggtggagaag tgcctagata
     2221 aatatttcca gcatctttgt gatgatctgg aggtatttgc tgctcatgct ggccgcaaga
     2281 ctgtgaagcc agaggacctg gagctgctga tgcggcggca gggcctggtc actgaccaag
     2341 tctcactgca cgtgctagtg gagcggcacc tgcccctgga gtaccggcag ctgctcatcc
     2401 cctgtgcata cagtggcaac tctgtcttcc ctgcccaggg atccgactac aaagaccatg
     2461 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     2521 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2581 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2641 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2701 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2761 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2821 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2881 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     2941 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3001 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3061 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3121 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3181 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     3241 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     3301 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     3361 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     3421 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3481 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3541 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     3601 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3661 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3721 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3781 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     3841 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3901 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3961 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4021 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4081 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4141 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4201 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4261 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4321 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4381 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4441 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4501 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4561 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4621 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4681 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4741 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4801 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4861 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4921 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4981 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5041 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     5101 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5161 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5221 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5281 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5341 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5401 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5461 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5521 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5581 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5641 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5701 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5761 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5821 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5881 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5941 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6001 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6061 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6241 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6481 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6541 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6601 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6661 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6721 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6781 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6841 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6901 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6961 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7021 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7081 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7141 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7201 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7261 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7321 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7381 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7441 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7501 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7561 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     7621 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     7681 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     7741 atgaagaatc tgcttagg
//
