LOCUS       sequence-72063-e        1034 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Golden Gate Compatible TALEN Construct for site specific 
            modification of genome, +63 C Terminus, Codon Optimized FokI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1034)
  TITLE     Bao Lab Golden Gate plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1034)
  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: 
            "Unpublished"
COMMENT     Sequence Label: TALEN_C63
FEATURES             Location/Qualifiers
     source          1..1034
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     7..26
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(73..93)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(73..91)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(112..128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(112..128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(125..147)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    136..152
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(160..166,167..184,185..190))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    205..226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             412..999
                     /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
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
ORIGIN
        1 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcctc
       61 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
      121 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
      181 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
      241 actgcccgct ttccaccggt cgtctccaac gaccatctgg tggcgttggc atgtcttggt
      301 ggacgacccg cgctcgatgc agtcaaaaag ggtctgcctc atgctcccgc attgatcaaa
      361 agaaccaacc ggcggattcc cgagagaact tcccatcgag tcgcgggatc ccagctggtg
      421 aagagcgagc tggaggagaa gaagtccgag ctgcggcaca agctgaagta cgtgccccac
      481 gagtacatcg agctgatcga gatcgccagg aacagcaccc aggaccgcat cctggagatg
      541 aaggtgatgg agttcttcat gaaggtgtac ggctacaggg gaaagcacct gggcggaagc
      601 agaaagcctg acggcgccat ctatacagtg ggcagcccca tcgattacgg cgtgatcgtg
      661 gacacaaagg cctacagcgg cggctacaat ctgcctatcg gccaggccga cgagatgcag
      721 agatacgtgg aggagaacca gacccggaat aagcacatca accccaacga gtggtggaag
      781 gtgtacccta gcagcgtgac cgagttcaag ttcctgttcg tgagcggcca cttcaagggc
      841 aactacaagg cccagctgac caggctgaac cacatcacca actgcaatgg cgccgtgctg
      901 agcgtggagg agctgctgat cggcggcgag atgatcaaag ccggcaccct gacactggag
      961 gaggtgcggc gcaagttcaa caacggcgag atcaacttct gataacttaa gtttaaaccg
     1021 ctgatcagcc tcga
//
