LOCUS       sequence-48821-e         843 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 843)
  AUTHORS   Miller JC, Holmes MC, Wang J, Guschin DY, Lee YL, Rupniewski I, 
            Beausejour CM, Waite AJ, Wang NS, Kim KA, Gregory PD, Pabo CO, Rebar
            EJ
  TITLE     An improved zinc-finger nuclease architecture for highly specific 
            genome editing.
  JOURNAL   Nat Biotechnol. 2007 Jul . 25(7):778-85.
  PUBMED    17603475
REFERENCE   2  (bases 1 to 843)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2007 Jul . 25(7):778-85."
COMMENT     Sequence Label: pCLR2070
FEATURES             Location/Qualifiers
     source          1..843
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     36..55
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        36..54
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             83..103
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             113..136
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             159..746
                     /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="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMER
                     YVEENQTRDKHLNPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     CDS             762..803
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             813..830
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
       61 caagctggct agcaccatgg gacctaagaa aaagaggaag gtggcggccg ctgactacaa
      121 ggatgacgac gataaatcta gatagtaagc ttggatccca actagtcaaa agtgaactgg
      181 aggagaagaa atctgaactt cgtcataaat tgaaatatgt gcctcatgaa tatattgaat
      241 taattgaaat tgccagaaat tccactcagg atagaattct tgaaatgaag gtaatggaat
      301 tttttatgaa agtttatgga tatagaggta aacatttggg tggatcaagg aaaccggacg
      361 gagcaattta tactgtcgga tctcctattg attacggtgt gatcgtggat actaaagctt
      421 atagcggagg ttataatctg ccaattggcc aagcagatga aatggagcga tatgtcgaag
      481 aaaatcaaac acgagacaaa catctcaacc ctaatgaatg gtggaaagtc tatccatctt
      541 ctgtaacgga atttaagttt ttatttgtga gtggtcactt taaaggaaac tacaaagctc
      601 agcttacacg attaaatcat atcactaatt gtaatggagc tgttcttagt gtagaagagc
      661 ttttaattgg tggagaaatg attaaagccg gcacattaac cttagaggaa gtgagacgga
      721 aatttaataa cggcgagata aacttttaag ggcccttcga aggtaagcct atccctaacc
      781 ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa
      841 ccc
//
