LOCUS       Exported                3707 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKK-platform DNA, complete sequence.
ACCESSION   AB781744
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3707
                     /organism="Cloning vector pKK-platform"
                     /mol_type="other DNA"
                     /db_xref="taxon:1291490"
     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             696..1421
                     /codon_start=1
                     /gene="ZFN"
                     /product="FLAG-NLS-partial ZF with EL mutated FokI nuclease
                     domain"
                     /label=ZFN
                     /protein_id="BAN13561.1"
                     /translation="MDYKDDDDKISPKKKRKVGSQKPFQCRICMRNFSCPYTKIHLRGS
                     QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNPTQDRILEMKVMEFFMKVYGYRGEH
                     LGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQRYVKENQTRNKHINP
                     NEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHKTNCNGAVLSVEELLIGGEMIKA
                     GTLTLEEVRRKFNNGEINF"
     gene            696..1421
                     /gene="ZFN"
                     /label=ZFN
     CDS             699..722
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             729..749
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             831..1418
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNPTQDRILEMKV
                     MEFFMKVYGYRGEHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVKENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHKTNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     promoter        complement(1622..1640)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1872..1976
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1977..2837
                     /codon_start=1
                     /gene="amp"
                     /product="beta-lactamase class A"
                     /label=amp
                     /protein_id="BAN13562.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            1977..2837
                     /gene="amp"
                     /label=amp
     rep_origin      3008..3596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"