LOCUS       sequence-474068-         778 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  bacterial expression of human TTR mutants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 778)
  AUTHORS   Sun X, Ferguson JA, Leach BI, Stanfield RL, Dyson HJ, Wright PE
  TITLE     Probing the Dissociation Pathway of a Kinetically Labile 
            Transthyretin Mutant.
  JOURNAL   J Am Chem Soc. 2024 Jan 10;146(1):532-542. doi: 
            10.1021/jacs.3c10083. Epub 2023 Dec 22.
  PUBMED    38134439
REFERENCE   2  (bases 1 to 778)
  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"; doi: 
            "10.1021/jacs.3c10083"; journalName: "J Am Chem Soc"; date: 
            "2024-01-10- 10"; volume: "146"; issue: "1"; pages: "532-542"
COMMENT     Sequence Label: His6-tev-G(TTR A25T)
FEATURES             Location/Qualifiers
     source          1..778
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     22..41
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     125..144
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        125..143
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    144..168
                     /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             183..205
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             218..235
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             266..286
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             719..736
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
       61 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
      121 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
      181 attttgttta actttaagaa ggagatatac catgggccat catcatcatc atcacagcgg
      241 cgaattcagc agcggcagca gcggcgaaaa cctgtatttt cagggtggcc ctacgggcac
      301 cggtgaatcc aagtgtcctc tgatggtcaa agttctagat gctgtccgag gcagtcctac
      361 catcaatgtg gccgtgcatg tgttcagaaa ggctgctgat gacacctggg agccatttgc
      421 ctctgggaaa accagtgagt ccggagagct gcatgggctc acaactgagg aggaatttgt
      481 agaagggata tacaaagtgg aaatagacac caaatcttac tggaaggcac ttggcatctc
      541 cccattccat gagcatgcag aggtggtatt cacagccaac gactccggcc cccgccgcta
      601 caccattgcc gccctgctga gcccctactc ctattccacc acggctgtcg tcaccaatcc
      661 caaggaataa taggatccga attcgagctc cgtcgacaag cttgcagccg cactcgagca
      721 ccaccaccac caccactgag aatccggctg ctaacaaagc ccgaaaggaa gcgatgtg
//
