LOCUS       sequence-72004-e        1075 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 1075)
  TITLE     Bao Lab Golden Gate plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1075)
  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            "Unpublished"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..1075
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     23..43
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        25..43
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(91..109)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(119..136)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(119..135)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(128..150)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(732..752)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(759..824)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(831..848)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(858..875)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             894..916
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    931..955
                     /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(956..974)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagga
       61 cctcgagggg gggcccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
      121 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
      181 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
      241 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcccgtc
      301 tccagggggg caccggtgag cgcattgcgc caggcgtgca ctgcctctac cgctgttact
      361 ccccctctct tcgcgatctt cagcagctgc ccggtgtcga gctggagcgg aggcccccta
      421 agctcacccg ccacagtcag cagcgcctca agtgctcgcg ctcccgacca ctgtttaccg
      481 acccctacaa ttgcctcgtg cgtggcttcg ggcagggccg caatcatatc ttggtatttg
      541 acagccaccg tcccaagcgc cgcagggtgc tgtgaaagcg cgacaatatg cgcatgagtg
      601 aagccatgcc ccacaagcgc ctcgtggtgt tgcgcgacgg tgctcctgac cttaggcttg
      661 attttctcct gttgctgttg cgaataaccg agtgtcctca agtccaccat aggtaccccg
      721 cggtgaatgc ccaccttcct cttcttcttg ggggccatct tgtcatcgtc atccttgtaa
      781 tcgatgtcat gatctttata atcaccgtca tggtctttgt agtccatatg gctgccgcgc
      841 ggcaccaggc cgctgctgtg atgatgatga tgatggctgc tgcccatggt atatctcctt
      901 cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca attcccctat
      961 agtgagtcgt attaatttcg cgggatcgag atctcgatcc tctacgccgg acgcatcgtg
     1021 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catca
//
