LOCUS       sequence-71895-e        1009 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 1009)
  TITLE     Bao Lab Golden Gate plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1009)
  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            "Unpublished"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..1009
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        3..21
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      41..50
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             50..115
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             122..142
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(299..874)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TGERIAPGVHCLYRCYSPSLRDLQQLPGVELERRPPKLTRHSQQRLKCSRSRPLFTDPY
                     NCLVRGFGQGRNHILVFDSHRPKRRRVL"
     primer_bind     724..746
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     738..755
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     739..755
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        765..783
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(831..851)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(831..849)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(870..886)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(870..886)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(883..905)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    894..910
                     /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(918..924,925..942,943..948))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    963..984
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 attaatacga ctcactatag ggagacccaa gctggctagc gccaccatgg actacaaaga
       61 ccatgacggt gattataaag atcatgacat cgattacaag gatgacgatg acaagatggc
      121 ccccaagaag aagaggaagg tgggcattca ccgcggggta cctatggtgg acttgaggac
      181 actcggttat tcgcaacagc aacaggagaa aatcaagcct aaggtcagga gcaccgtcgc
      241 gcaacaccac gaggcgcttg tggggcatgg cttcactcat gcgcatattg tcgcgctttc
      301 acagcaccct gcggcgcttg ggacggtggc tgtcaaatac caagatatga ttgcggccct
      361 gcccgaagcc acgcacgagg caattgtagg ggtcggtaaa cagtggtcgg gagcgcgagc
      421 acttgaggcg ctgctgactg tggcgggtga gcttaggggg cctccgctcc agctcgacac
      481 cgggcagctg ctgaagatcg cgaagagagg gggagtaaca gcggtagagg cagtgcacgc
      541 ctggcgcaat gcgctcaccg gtgcccccct ggagacgggc gccgctacag ggcgcgtccc
      601 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      661 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      721 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
      781 agggcgaatt gggtaccggg ccccccctcg aggtcctcca gcttttgttc cctttagtga
      841 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
      901 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
      961 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgctt
//
