LOCUS       sequence-72060-e        1068 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 1068)
  TITLE     Bao Lab Golden Gate plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1068)
  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            "Unpublished"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..1068
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(13..600)
                     /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    651..672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(687..692,693..710,711..717)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    725..741
                     /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     730..752
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     749..765
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     749..765
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             761..1066
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQ"
     primer_bind     784..804
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        786..804
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(851..870)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(852..870)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(880..897)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(880..896)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(889..911)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cttaagttat cagaagttga tctcgccgtt gttgaacttg cgccgcacct cctccagtgt
       61 cagggtgccg gctttgatca tctcgccgcc gatcagcagc tcctccacgc tcagcacggc
      121 gccattgcag ttggtgatgt ggttcagcct ggtcagctgg gccttgtagt tgcccttgaa
      181 gtggccgctc acgaacagga acttgaactc ggtcacgctg ctagggtaca ccttccacca
      241 ctcgttgggg ttgatgtgct tattccgggt ctggttctcc tccacgtatc tctgcatctc
      301 gtcggcctgg ccgataggca gattgtagcc gccgctgtag gcctttgtgt ccacgatcac
      361 gccgtaatcg atggggctgc ccactgtata gatggcgccg tcaggctttc tgcttccgcc
      421 caggtgcttt cccctgtagc cgtacacctt catgaagaac tccatcacct tcatctccag
      481 gatgcggtcc tgggtgctgt tcctggcgat ctcgatcagc tcgatgtact cgtggggcac
      541 gtacttcagc ttgtgccgca gctcggactt cttctcctcc agctcgctct tcaccagctg
      601 ggatccgttg gagacgaccg gtggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
      661 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
      721 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
      781 cgcgcaatta accctcacta aagggaacaa aagctggagg acctcgaggg ggggcccggt
      841 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
      901 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
      961 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     1021 cctgaatggc gaatgggacg cgccctgtag cggcgcccgt ctccaggg
//
