LOCUS       sequence-72062-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  JOURNAL   Unpublished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            "Unpublished"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"
     primer_bind     4..24
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6..24
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(72..90)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(100..117)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(100..116)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(109..131)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(713..733)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(740..805)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     RBS             818..840
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    855..879
                     /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(880..898)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(982..1001)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cgcgcaatta accctcacta aagggaacaa aagctggagg acctcgaggg ggggcccggt
       61 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
      121 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
      181 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
      241 cctgaatggc gaatgggacg cgccctgtag cggcgcccgt ctccaggggg gcaccggtga
      301 gcgcattgcg ccaggcgtgc actgcctcta ccgctgttac tccccctctc ttcgcgatct
      361 tcagcagctg cccggtgtcg agctggagcg gaggccccct aagctcaccc gccacagtca
      421 gcagcgcctc aagtgctcgc gctcccgacc actgtttacc gacccctaca attgcctcgt
      481 gcgtggcttc gggcagggcc gcaatcatat cttggtattt gacagccacc gtcccaagcg
      541 ccgcagggtg ctgtgaaagc gcgacaatat gcgcatgagt gaagccatgc cccacaagcg
      601 cctcgtggtg ttgcgcgacg gtgctcctga ccttaggctt gattttctcc tgttgctgtt
      661 gcgaataacc gagtgtcctc aagtccacca taggtacccc gcggtgaatg cccaccttcc
      721 tcttcttctt gggggccatc ttgtcatcgt catccttgta atcgatgtca tgatctttat
      781 aatcaccgtc atggtctttg tagtccgcca tggtatatct ccttcttaaa gttaaacaaa
      841 attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag tcgtattaat
      901 ttcgcgggat cgagatctcg atcctctacg ccggacgcat cgtggccggc atcaccggcg
      961 ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc
     1021 gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggt
//
