LOCUS       sequence-72007-e        1037 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  for e. coli expression, Golden Gate Compatible TALEN incorporated 
            with N Terminal 6xHIS Tag, Codon Optimized FokI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1037)
  TITLE     Bao Lab Golden Gate plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1037)
  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            "Unpublished"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-HIS-Sangamo
FEATURES             Location/Qualifiers
     source          1..1037
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(39..56)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(100..687)
                     /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    873..894
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(909..914,915..932,933..939)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    947..963
                     /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     952..974
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     971..987
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     971..987
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1006..1026
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1008..1026
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tcagcttcct ttcgggcttt gttagcagcc ggatctcagt ggtggtggtg gtggtgctcg
       61 agtgcggccg caagcttgtc gacggagctc gaattctcag aagttgatct cgccgttgtt
      121 gaacttgcgc cgcacctcct ccagtgtcag ggtgccggct ttgatcatct cgccgccgat
      181 cagcagctcc tccacgctca gcacggcgcc attgcagttg gtgatgtggt tcagcctggt
      241 cagctgggcc ttgtagttgc ccttgaagtg gccgctcacg aacaggaact tgaactcggt
      301 cacgctgcta gggtacacct tccaccactc gttggggttg atgtgcttat tccgggtctg
      361 gttctcctcc acgtatctct gcatctcgtc ggcctggccg ataggcagat tgtagccgcc
      421 gctgtaggcc tttgtgtcca cgatcacgcc gtaatcgatg gggctgccca ctgtatagat
      481 ggcgccgtca ggctttctgc ttccgcccag gtgctttccc ctgtagccgt acaccttcat
      541 gaagaactcc atcaccttca tctccaggat gcggtcctgg gtgctgttcc tggcgatctc
      601 gatcagctcg atgtactcgt ggggcacgta cttcagcttg tgccgcagct cggacttctt
      661 ctcctccagc tcgctcttca ccagctggga tcccgcgact cgatgggaag ttctctcggg
      721 aatccgccgg ttggttcttt tgatcaatgc gggagcatga ggcagaccct ttttgactgc
      781 atcgagcgcg ggtcgtccac caagacatgc caacgccacc agatggtcgt tggagacgac
      841 cggtggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
      901 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
      961 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     1021 taaagggaac aaaagct
//
