LOCUS       sequence-41155-e         880 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 880)
  AUTHORS   Foley JE, Maeder ML, Pearlberg J, Joung JK, Peterson RT, Yeh JR
  TITLE     Targeted mutagenesis in zebrafish using customized zinc-finger 
            nucleases.
  JOURNAL   Nat Protoc. 2009. 4(12):1855-67.
  PUBMED    20010934
REFERENCE   2  (bases 1 to 880)
  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 Protoc.
            2009. 4(12):1855-67."
COMMENT     Sequence Label: pMLM290
FEATURES             Location/Qualifiers
     source          1..880
                     /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
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVKENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHKTNCNGAVL
                     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"
     primer_bind     complement(853..870)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
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 aatgcaacga tatgtcaaag
      481 aaaatcaaac acgaaacaaa catatcaacc ctaatgaatg gtggaaagtc tatccatctt
      541 ctgtaacgga atttaagttt ttatttgtga gtggtcactt taaaggaaac tacaaagctc
      601 agcttacacg attaaatcat aagactaatt 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 cccgctgatc agcctcgact gtgccttcta gttgccagcc
//
