LOCUS       sequence-72061-e        1085 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  for e. coli expression, Golden Gate Compatible TALEN incorporated 
            with C Terminal 6xHIS Tag, Codon Optimized FokI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1085)
  TITLE     Bao Lab Golden Gate plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1085)
  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            "Unpublished"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: pET-Sangamo-HIS
FEATURES             Location/Qualifiers
     source          1..1085
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     45..67
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     59..76
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     60..76
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     86..105
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        86..104
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(152..172)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(152..170)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(191..207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(191..207)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(204..226)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    215..231
                     /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(239..245,246..263,264..269))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    284..305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             491..1078
                     /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 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
       61 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      121 gccccccctc gaggtcctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
      181 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      241 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      301 cattaattgc gttgcgctca ctgcccgctt tccaccggtc gtctccaacg accatctggt
      361 ggcgttggca tgtcttggtg gacgacccgc gctcgatgca gtcaaaaagg gtctgcctca
      421 tgctcccgca ttgatcaaaa gaaccaaccg gcggattccc gagagaactt cccatcgagt
      481 cgcgggatcc cagctggtga agagcgagct ggaggagaag aagtccgagc tgcggcacaa
      541 gctgaagtac gtgccccacg agtacatcga gctgatcgag atcgccagga acagcaccca
      601 ggaccgcatc ctggagatga aggtgatgga gttcttcatg aaggtgtacg gctacagggg
      661 aaagcacctg ggcggaagca gaaagcctga cggcgccatc tatacagtgg gcagccccat
      721 cgattacggc gtgatcgtgg acacaaaggc ctacagcggc ggctacaatc tgcctatcgg
      781 ccaggccgac gagatgcaga gatacgtgga ggagaaccag acccggaata agcacatcaa
      841 ccccaacgag tggtggaagg tgtaccctag cagcgtgacc gagttcaagt tcctgttcgt
      901 gagcggccac ttcaagggca actacaaggc ccagctgacc aggctgaacc acatcaccaa
      961 ctgcaatggc gccgtgctga gcgtggagga gctgctgatc ggcggcgaga tgatcaaagc
     1021 cggcaccctg acactggagg aggtgcggcg caagttcaac aacggcgaga tcaacttcgc
     1081 ggccg
//
