LOCUS       V021692                 9104 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021692
VERSION     V021692
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..3751
                     /label="CDCA2(NM_152562)"
                     /note="CDCA2(NM_152562)"
                     /gene="CDCA2"
     CDS             3791..3856
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3880..4468
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4534..4582
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4784..5212
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5226..5555
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5622..6416
                     /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    6592..6713
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6762..6778)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6786..6802
                     /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(6810..6840)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6855..6876
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7164..7752)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7923..8783)
                     /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(8784..8888)
                     /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 ccatggatgc caattcaaaa gacaagcccc ctgaaaccaa
      721 ggagtctgca atgaataatg ctggaaatgc ctctttcatt ttgggaactg ggaagattgt
      781 gactcctcag aagcatgccg aattacctcc taatccttgc acaccagata cttttaaatc
      841 acctttgaac ttttccacag taaccgtaga gcaattggga attacacctg aaagctttgt
      901 taggaactct gcaggaaagt catcatccta ccttaaaaaa tgtagacgac gttctgcagt
      961 cggtgctcgg ggctctcctg aaacaaacca tctgattcgt ttcattgctc ggcagcaaaa
     1021 tataaagaat gctaggaaat ctcctttggc acaagattct ccttcccagg gcagccctgc
     1081 actgtatcga aatgttaaca ctttaagaga acgaatatca gccttccagt cagcttttca
     1141 ctccataaag gaaaacgaga aaatgaccgg ctgtctggaa ttctcagagg caggaaaaga
     1201 gtccgagatg acagacttga ccagaaagga aggtctcagc gcttgccagc agtctgggtt
     1261 ccctgcagtg ttgtcctcca aacgtcggag aatatcctat cagagagact ctgatgaaaa
     1321 tctgacggat gctgaaggaa aagtaattgg tctccagata ttcaatattg atacagacag
     1381 agcatgtgca gttgaaactt ctgtagatct ttctgagata tcatctaaac ttggttcaac
     1441 acagtctgga tttttagttg aagagtctct tcccctttca gagctcacag agacttcaaa
     1501 tgcactaaag gttgctgact gtgtagtggg caaaggatca agtgatgccg tttcgcctga
     1561 cacgttcaca gcagaagtga gctcagacgc agtccctgat gtcaggtcac cagctactcc
     1621 agcctgcagg agggaccttc ccacccccaa gacctttgta cttcgttctg tactgaagaa
     1681 accctctgtt aagatgtgtc tagagagctt acaggaacac tgtaacaacc tctatgatga
     1741 tgatgggact catccgagct taatctcaaa tctcccaaac tgttgcaaag agaaagaagc
     1801 agaagatgaa gaaaattttg aagcacctgc ctttctaaat atgaggaaga ggaagagagt
     1861 tacttttgga gaggacttaa gcccggaagt gtttgatgaa tctttgccag caaatactcc
     1921 attgcgtaaa ggaggaacac ctgtttgtaa aaaagacttc agtggtctca gttccctgct
     1981 gcttgagcag tcacctgttc ctgagccatt acctcaacca gattttgatg acaaggggga
     2041 gaatcttgaa aacatagaac cacttcaagt atcatttgcc gttctcagtt ctcctaataa
     2101 atcatcaatc tctgagaccc tttcaggcac tgataccttt agttcttcaa ataaccatga
     2161 gaaaatatcc tctcctaaag ttggtagaat aacaaggact tctaacagaa gaaatcaatt
     2221 ggtcagtgtt gtagaagaga gtgtttgcaa cttattgaat acagaagttc agccttgtaa
     2281 agaaaagaaa attaatagga ggaagtctca agaaacaaag tgtacaaaga gagcacttcc
     2341 taagaagagt caggttttaa aaagttgcag aaagaagaaa ggaaagggaa agaaaagtgt
     2401 tcagaaatct ttatatgggg aaagagacat tgcttctaag aagcccctcc tcagtcctat
     2461 tcccgagctg cctgaagtcc ctgagatgac accttccatt ccgagcatcc gaagactggg
     2521 ttcaggttat ttcagttcaa atggcaaact ggaagaagtg aagactccta aaaatccagt
     2581 gaaaagaaag gatcttttgc gtcatgaccc agatttgcat atgcatcaag gctatgataa
     2641 atatgatgtc tctgaattct gctcttatat aaaaagttcc tcatcgcttg gcaatgctac
     2701 ttctgatgaa gatccaaata caaatataat gaacattaat gaaaataaaa atattccaaa
     2761 agcaaaaaat aagtcagaaa gtgaaaatga accaaaagct ggaactgaca gtcctgtttc
     2821 ttgtgcttct gtaactgaag aacgtgtggc atcagatagt cccaaacctg ctctgaccct
     2881 gcagcagggt caagaatttt ctgctggtgg tcaaaatgca gaaaaccttt gtcagttctt
     2941 taaaatttca ccagatttaa acataaagtg tgaaagaaag gatgacttct taggagctgc
     3001 agaaggaaaa ctgcaatgca atcgtttaat gcctaattca caaaaagact gtcattgttt
     3061 aggagatgtc ttaattgaaa atacgaaaga atctaaaagc cagagtgagg atttgggaag
     3121 aaaacccatg gaaagtagca gtgttgtgag ttgcagagac aggaaagata gaagacgttc
     3181 catgtgttat tctgatggtc gaagtttaca tttggaaaaa aatggaaatc acacaccatc
     3241 ctccagtgtg ggcagctctg tagaaattag tttagaaaat tctgaactgt ttaaagattt
     3301 gtctgatgcc attgagcaaa cctttcagag gagaaatagt gaaaccaaag tgcgacgtag
     3361 cacgaggcta cagaaggatt tagaaaacga aggtcttgta tggatttcac ttccacttcc
     3421 ttccacttcc caaaaagcca aaagaagaac aatatgtaca tttgacagca gtggatttga
     3481 aagtatgtct cccataaaag aaactgtgtc ctccagacaa aaaccgcaga tggcacctcc
     3541 cgtctcagat ccagaaaaca gccagggccc tgctgctggt tcttccgatg aacctggtaa
     3601 gaggaggaag agcttttgta tatctacact tgcaaatact aaagccactt cccagttcaa
     3661 aggctaccgg agaagatcct ctcttaatgg gaagggagag agctctctga ctgccttgga
     3721 aaggattgaa cataatggag aaagaaagca ggaattctgc agatatccag cacagtggcg
     3781 gccgctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     3841 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa
     3901 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3961 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     4021 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     4081 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     4141 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4201 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4261 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     4321 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4381 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4441 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga
     4501 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     4561 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4621 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4681 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4741 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     4801 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4861 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4921 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     4981 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5041 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5101 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     5161 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     5221 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5281 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     5341 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5401 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5461 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     5521 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     5581 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5641 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5701 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5761 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     5821 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5881 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5941 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     6001 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     6061 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6121 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     6181 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6241 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6301 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6361 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6421 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6481 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6541 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     6601 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6661 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6721 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6781 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6841 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6901 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6961 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7021 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7081 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7141 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7201 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7261 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7321 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     7381 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7441 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7501 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7561 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7621 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7681 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7741 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7801 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7861 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7921 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7981 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     8041 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8101 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8161 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8221 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8281 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8341 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8401 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8461 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8521 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8581 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8641 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8701 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8761 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8821 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8881 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     8941 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     9001 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     9061 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
