LOCUS       Exported                1056 bp ds-DNA     linear   SYN 19-MAY-2021
DEFINITION  for e. coli expression, Golden Gate Compatible TALEN incorporated 
            with N Terminal 6xHIS Tag, Codon Optimized FokI.
ACCESSION   .
VERSION     .
KEYWORDS    pET-HIS-Sangamo
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1056)
  TITLE     Bao Lab Golden Gate plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1056)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 19, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..1056
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(3..79)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     285..304
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     388..407
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        388..406
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    407..431
                     /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)."
     RBS             446..468
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             487..504
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             514..531
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             538..603
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             610..630
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
       61 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
      121 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
      181 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
      241 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
      301 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      361 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
      421 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata taccatgggc
      481 agcagccatc atcatcatca tcacagcagc ggcctggtgc cgcgcggcag ccatatggac
      541 tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac
      601 aagatggccc ccaagaagaa gaggaaggtg ggcattcacc gcggggtacc tatggtggac
      661 ttgaggacac tcggttattc gcaacagcaa caggagaaaa tcaagcctaa ggtcaggagc
      721 accgtcgcgc aacaccacga ggcgcttgtg gggcatggct tcactcatgc gcatattgtc
      781 gcgctttcac agcaccctgc ggcgcttggg acggtggctg tcaaatacca agatatgatt
      841 gcggccctgc ccgaagccac gcacgaggca attgtagggg tcggtaaaca gtggtcggga
      901 gcgcgagcac ttgaggcgct gctgactgtg gcgggtgagc ttagggggcc tccgctccag
      961 ctcgacaccg ggcagctgct gaagatcgcg aagagagggg gagtaacagc ggtagaggca
     1021 gtgcacgcct ggcgcaatgc gctcaccggt gccccc
//
