LOCUS       sequence-72012-e        1063 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 1063)
  TITLE     Bao Lab Golden Gate plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1063)
  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            "Unpublished"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..1063
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(39..56)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(72..659)
                     /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"
     protein_bind    845..866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(881..886,887..904,905..911)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    919..935
                     /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)."
     primer_bind     924..946
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     943..959
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     943..959
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     978..998
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        980..998
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tcagcttcct ttcgggcttt gttagcagcc ggatctcagt ggtggtggtg gtggtgctcg
       61 agtgcggccg cgaagttgat ctcgccgttg ttgaacttgc gccgcacctc ctccagtgtc
      121 agggtgccgg ctttgatcat ctcgccgccg atcagcagct cctccacgct cagcacggcg
      181 ccattgcagt tggtgatgtg gttcagcctg gtcagctggg ccttgtagtt gcccttgaag
      241 tggccgctca cgaacaggaa cttgaactcg gtcacgctgc tagggtacac cttccaccac
      301 tcgttggggt tgatgtgctt attccgggtc tggttctcct ccacgtatct ctgcatctcg
      361 tcggcctggc cgataggcag attgtagccg ccgctgtagg cctttgtgtc cacgatcacg
      421 ccgtaatcga tggggctgcc cactgtatag atggcgccgt caggctttct gcttccgccc
      481 aggtgctttc ccctgtagcc gtacaccttc atgaagaact ccatcacctt catctccagg
      541 atgcggtcct gggtgctgtt cctggcgatc tcgatcagct cgatgtactc gtggggcacg
      601 tacttcagct tgtgccgcag ctcggacttc ttctcctcca gctcgctctt caccagctgg
      661 gatcccgcga ctcgatggga agttctctcg ggaatccgcc ggttggttct tttgatcaat
      721 gcgggagcat gaggcagacc ctttttgact gcatcgagcg cgggtcgtcc accaagacat
      781 gccaacgcca ccagatggtc gttggagacg accggtggaa agcgggcagt gagcgcaacg
      841 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
      901 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
      961 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gaggacctcg
     1021 agggggggcc cggtacccaa ttcgccctat agtgagtcgt att
//
