LOCUS       V018585                 8183 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018585
VERSION     V018585
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..2830
                     /label="Kif2a(NM_008442)"
                     /note="Kif2a(NM_008442)"
                     /gene="Kif2a"
     CDS             2870..2935
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2959..3547
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3613..3661
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3863..4291
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4305..4634
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4701..5495
                     /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    5671..5792
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5841..5857)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5865..5881
                     /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(5889..5919)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5934..5955
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6243..6831)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7002..7862)
                     /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(7863..7967)
                     /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 ccatggtaac atctttaaat gaagataatg aaagtgtaac
      721 tgttgagtgg atagaaaatg gagatacgaa aggcaaagag attgacttgg agagcatctt
      781 ttcacttaac cctgaccttg tacctgatga agatattgag cccagtccag aactacctcc
      841 accctcgtca tcctcaaaag ttaacaaaat tgtaaagaac cggcggactg tggcagctgt
      901 taagaatgac cctcccccga gagataatag agtggttggt tcagcacgcg cacggcctag
      961 tcagcttcct gagcaatcgt cttctgcaca acagaatggt agcgtttcag atatatctcc
     1021 agttcaagct gcaaaaaagg agtttggccc tccttcacgt agaaaatcca attgtgtgaa
     1081 agaagtagaa aaattgcaag aaaaacgaga aaaaaggaga ttgcaacagc aagaacttag
     1141 agaaaaaaga gcccaggatg ttgatgctac aaatccaaat tacgaaatta tgtgtatgat
     1201 cagagacttc agagggagct tggattacag acccctaaca acagcagatc ctattgatga
     1261 acataggata tgtgtttgtg taagaaaacg accactcaat aaaaaagaaa ctcaaatgaa
     1321 agatcttgat gtgatcacaa tccctagtaa agatgttgtg atggtccacg aacccaaaca
     1381 gaaagtagac ttaacaaggt acttagaaaa tcagacattt cggtttgatt acgccttcga
     1441 tgactcagct cctaatgaaa tggtttacag gtttactgct aggccactag tggaaacaat
     1501 atttgaaagg ggcatggcta cgtgctttgc ttatgggcag actggaagtg ggaaaactca
     1561 tactatgggt ggtgactttt caggaaagaa ccaggattgt tctaagggaa tttatgcatt
     1621 agcagctcgc gatgtctttt taatgctgaa gaagccaaac tataagaaac tagaacttca
     1681 agtatatgca acattttttg aaatttatag tggaaaggtg tttgacttac taaataggaa
     1741 aacaaaatta agagtactag aagatgggaa gcagcaggtc caagtggtgg ggctgcagga
     1801 gcgggaggtg aagtgtgtgg aggacgtgct gaagctcatc gacataggca acagctgcag
     1861 aacatctggc caaacatctg caaatgcaca ttcatctcgg agccatgcag tgttccagat
     1921 cattctaaga aggaagggga aactacatgg taaattttct ctcattgacc tggctgggaa
     1981 cgaaagagga gctgatacat ccagtgcaga caggcaaacc aggcttgaag gtgctgaaat
     2041 taataaaagc cttttagcac tcaaggagtg catccgagcc ttaggtagaa acaaacccca
     2101 tacgcctttc cgagcgagca aactcactca ggtgctgaga gattccttca taggtgaaaa
     2161 ttctcgtacc tgcatgattg ccacaatctc tccaggaatg gcatcctgtg aaaatactct
     2221 taatacgtta agatatgcaa atagagtaaa ggagtttgga attagtcctt cagacattcc
     2281 cttctcccag ggtggtggca gtcgccctga tctctctcct tcttacgatt atgacgactt
     2341 ttctccttca attaccaggg tgaaagaatt gactgtaaat ccagctgctg ctggcgatgt
     2401 ccatccaata atgcaccatc caccgagcca gattgatgac ttagagacac agtggggtgt
     2461 ggggagttct cctcagagag atgatctaaa acttctctgt gaacaaaatg aagaagaagt
     2521 ttctccacag ctgttcactt tccatgaagc tgtctcccag atggtagaga tggaagagca
     2581 ggttgtggaa gatcataggg cagtgttcca ggaatccatt cgctggatag aagatgaaaa
     2641 ggctctccta gagatgactg aagaagtaga ctatgatgta gactcatatg ctacacaact
     2701 tgaagctatt cttgagcaaa aaatagacat tttaactgag ttacgagata aagtgaaatc
     2761 ttttcgcgca gcgctacaag aagaagagca agcaagcaag caaattaacc ccaagaggcc
     2821 ccgagccctc gaattctgca gatatccagc acagtggcgg ccgctcgagg actacaaaga
     2881 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg
     2941 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3001 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3061 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3121 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3181 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3241 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3301 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     3361 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3421 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3481 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3541 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3601 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     3661 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     3721 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3781 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     3841 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3901 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3961 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     4021 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4081 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4141 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4201 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     4261 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4321 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4381 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4441 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4501 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4561 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4621 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4681 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4741 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4801 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4861 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4921 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4981 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5041 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5101 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5161 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5221 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5281 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5341 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5401 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5461 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     5521 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5581 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5641 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5701 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5761 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5821 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5881 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5941 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6001 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6061 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6121 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6181 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6241 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6301 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6361 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6421 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6481 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6541 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6601 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6661 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6721 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6781 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6841 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6901 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6961 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7021 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7081 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7141 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7201 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7261 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7321 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7381 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7441 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7501 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7561 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7621 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7681 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7741 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7801 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7861 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7921 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7981 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca
     8041 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     8101 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     8161 attgcatgaa gaatctgctt agg
//
