LOCUS       sequence-33559-e         939 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 939)
  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 939)
  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: pMLM800
FEATURES             Location/Qualifiers
     source          1..939
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(2..19)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(29..70)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(86..670)
                     /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="LVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKVM
                     EFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQRY
                     VKENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHKTNCNGAVLS
                     VEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     CDS             complement(698..721)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(731..751)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(779..798)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(780..798)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(843..894)
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     primer_bind     complement(872..892)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 catggtgatg gtgatgatga ccggtacgcg tagaatcgag accgaggaga gggttaggga
       61 taggcttacc ttcgaagggc ccttaaaagt ttatctcgcc gttattaaat ttccgtctca
      121 cttcctctaa ggttaatgtg ccggctttaa tcatttctcc accaattaaa agctcttcta
      181 cactaagaac agctccatta caattagtct tatgatttaa tcgtgtaagc tgagctttgt
      241 agtttccttt aaagtgacca ctcacaaata aaaacttaaa ttccgttaca gaagatggat
      301 agactttcca ccattcatta gggttgatat gtttgtttcg tgtttgattt tctttgacat
      361 atcgttgcat ttcatctgct tggccaattg gcagattata acctccgcta taagctttag
      421 tatccacgat cacaccgtaa tcaataggag atccgacagt ataaattgct ccgtccggtt
      481 tccttgatcc acccaaatgt ttacctctat atccataaac tttcataaaa aattccatta
      541 ccttcatttc aagaattcta tcctgagtgg aatttctggc aatttcaatt aattcaatat
      601 attcatgagg cacatatttc aatttatgac gaagttcaga tttcttctcc tccagttcac
      661 ttttgactag ggctcgagcg gccgcatatt ttctagattt atcgtcgtca tccttgtagt
      721 cagcggcagc caccttcctc tttttcttag gtcccatggt gctagccagc ttgggtctcc
      781 ctatagtgag tcgtattaat ttcgataagc cagtaagcag tgggttctct agttagccag
      841 agagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg
      901 gcggagttgt tacgacattt tggaaagtcc cgttgattt
//
