LOCUS       sequence-72058-e        1064 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Golden Gate Compatible TALEN Construct site specific modification of
            genome, shortened C Terminus, Codon Optimized FokI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1064)
  TITLE     Bao Lab Golden Gate plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1064)
  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            "Unpublished"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_C18
FEATURES             Location/Qualifiers
     source          1..1064
                     /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             277..864
                     /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"
     primer_bind     complement(894..911)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    900..1011
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
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 ggatcccagc tggtgaagag cgagctggag
      301 gagaagaagt ccgagctgcg gcacaagctg aagtacgtgc cccacgagta catcgagctg
      361 atcgagatcg ccaggaacag cacccaggac cgcatcctgg agatgaaggt gatggagttc
      421 ttcatgaagg tgtacggcta caggggaaag cacctgggcg gaagcagaaa gcctgacggc
      481 gccatctata cagtgggcag ccccatcgat tacggcgtga tcgtggacac aaaggcctac
      541 agcggcggct acaatctgcc tatcggccag gccgacgaga tgcagagata cgtggaggag
      601 aaccagaccc ggaataagca catcaacccc aacgagtggt ggaaggtgta ccctagcagc
      661 gtgaccgagt tcaagttcct gttcgtgagc ggccacttca agggcaacta caaggcccag
      721 ctgaccaggc tgaaccacat caccaactgc aatggcgccg tgctgagcgt ggaggagctg
      781 ctgatcggcg gcgagatgat caaagccggc accctgacac tggaggaggt gcggcgcaag
      841 ttcaacaacg gcgagatcaa cttctgataa cttaagttta aaccgctgat cagcctcgac
      901 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
      961 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     1021 gagtaggtgt cattctattc tggggggtgg ggtggggcag gaca
//
