LOCUS       sequence-33560-e         896 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 896)
  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 896)
  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: pMLM802
FEATURES             Location/Qualifiers
     source          1..896
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(24..65)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(81..665)
                     /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
                     EFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMERY
                     VEENQTRNKHLNPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVLS
                     VEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     CDS             complement(693..716)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(726..746)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(774..793)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(775..793)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(838..889)
                     /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(867..887)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 tgatggtgat gatgaccggt acgcgtagaa tcgagaccga ggagagggtt agggataggc
       61 ttaccttcga agggccctta aaagtttatc tcgccgttat taaatttccg tctcacttcc
      121 tctaaggtta atgtgccggc tttaatcatt tctccaccaa ttaaaagctc ttctacacta
      181 agaacagctc cattacaatt agtgatatga tttaatcgtg taagctgagc tttgtagttt
      241 cctttaaagt gaccactcac aaataaaaac ttaaattccg ttacagaaga tggatagact
      301 ttccaccatt cattagggtt gagatgtttg tttcgtgttt gattttcttc gacatatcgc
      361 tccatttcat ctgcttggcc aattggcaga ttataacctc cgctataagc tttagtatcc
      421 acgatcacac cgtaatcaat aggagatccg acagtataaa ttgctccgtc cggtttcctt
      481 gatccaccca aatgtttacc tctatatcca taaactttca taaaaaattc cattaccttc
      541 atttcaagaa ttctatcctg agtggaattt ctggcaattt caattaattc aatatattca
      601 tgaggcacat atttcaattt atgacgaagt tcagatttct tctcctccag ttcacttttg
      661 actagggctc gagcggccgc atattttcta gatttatcgt cgtcatcctt gtagtcagcg
      721 gcagccacct tcctcttttt cttaggtccc atggtgctag ccagcttggg tctccctata
      781 gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta gccagagagc
      841 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atgggg
//
