LOCUS       40924_26794        3707 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKK-platform DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3707)
  AUTHORS   Fujii W, Kano K, Sugiura K, Naito K.
  TITLE     Repeatable Construction Method for Engineered Zinc Finger Nuclease 
            Based on Overlap Extension PCR and TA-cloning
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3707)
  AUTHORS   Fujii W, Kano K, Sugiura K, Naito K.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-FEB-2013) Contact:Wataru Fujii The University of 
            Tokyo, Graduate School of Agricultural and Life Sciences; Yayoi 
            1-1-1-7A-303, Bunkyo-ku, Tokyo 113-8657, Japan URL 
            :http://www.vm.a.u-tokyo.ac.jp/iden/
REFERENCE   3  (bases 1 to 3707)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3707)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-FEB-2013) Contact:Wataru Fujii The University of Tokyo, Graduate
            School of Agricultural and Life Sciences"; volume: " Yayoi 
            1-1-1-7A-303, Bunkyo-ku, Tokyo 113-8657, Japan URL :http"; pages: 
            "//www.vm.a.u-tokyo.ac.jp/iden"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             699..722
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             729..749
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             831..1418
                     /codon_start=1
                     /label=FokI cleavage domain
                     /note="nonspecific DNA cleavage domain of the FokI
                     endonuclease (Li et al., 1992)"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNPTQDRILEMKV
                     MEFFMKVYGYRGEHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVKENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHKTNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     promoter        complement(1622..1640)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1872..1976
                     /label=AmpR promoter
     CDS             1977..2834
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3008..3596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
      661 gcggtggcgg ccgctctagc ccgggcggat ccaccatgga ttacaaggat gacgacgata
      721 agatctcccc caagaagaag aggaaggtgg gatcacagaa gcccttccag tgccgcatct
      781 gcatgcgcaa cttcagctgc ccgtatacca agatccacct gcggggatcc caactagtca
      841 aaagtgaact ggaggagaag aaatctgaac ttcgtcataa attgaaatat gtgcctcatg
      901 aatatattga attaattgaa attgccagaa atcccactca ggatagaatt cttgaaatga
      961 aggtaatgga attttttatg aaagtttatg gatatagagg tgaacatttg ggtggatcaa
     1021 ggaaaccgga cggagcaatt tatactgtcg gatctcctat tgattacggt gtgatcgtgg
     1081 atactaaagc ttatagcgga ggttataatc tgccaattgg ccaagcagat gaaatgcaac
     1141 gatatgtcaa agaaaatcaa acacgaaaca aacatatcaa ccctaatgaa tggtggaaag
     1201 tctatccatc ttctgtaacg gaatttaagt ttttatttgt gagtggtcac tttaaaggaa
     1261 actacaaagc tcagcttaca cgattaaatc ataagactaa ttgtaatgga gctgttctta
     1321 gtgtagaaga gcttttaatt ggtggagaaa tgattaaagc cggcacatta accttagagg
     1381 aagtgagacg gaaatttaat aacggcgaga taaacttttg aggtacctat gggcaccaaa
     1441 gaacctgtaa acgttatctt tttaaattga atgtgcacaa ataaaagttt ggaaaagaaa
     1501 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1561 aaaaaaaaaa aaaaaaaaaa aaaaaaaccc tcgaggtacc aggtaagtgt acccaattcg
     1621 ccctatagtg agtcgtatta caattcactc gatcgccctt cccaacagtt gcgcagcctg
     1681 aatggcgaat ggagatccaa tttttaagtg tataatgtgt taaactactg attctaattg
     1741 tttgtgtatt ttagattcac agtcccaagg ctcatttcag gcagggcctc gtgatacgcc
     1801 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     1861 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1921 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1981 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2041 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2101 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2161 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2221 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2281 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2341 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2401 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2461 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2521 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2581 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2641 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2701 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2761 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2821 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     2881 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     2941 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3001 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3061 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3121 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttgt
     3661 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgcc
//
