LOCUS       sequence-146074-         897 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses a double mutant truncation of Hook3 in bacteria for 
            purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 897)
  AUTHORS   Schroeder CM, Vale RD
  TITLE     Assembly and activation of dynein-dynactin by the cargo adaptor 
            protein Hook3.
  JOURNAL   J Cell Biol. 2016 Aug 1;214(3):309-18. doi: 10.1083/jcb.201604002.
  PUBMED    27482052
REFERENCE   2  (bases 1 to 897)
  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: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2016-08-1- 1"; volume: "214"; 
            issue: "3"; pages: "309-18"
COMMENT     Sequence Label: pET28a-Hook3 D102A, E108A
FEATURES             Location/Qualifiers
     source          1..897
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     64..83
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        64..82
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    83..107
                     /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             122..144
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             162..179
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             189..206
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             210..242
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(258..281,282..317,318..341)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(378..406)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
       61 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
      121 ttttgtttaa ctttaagaag gagatataca tgggcagcag ccatcatcat catcatcaca
      181 gcagcggcct ggtgccgcgc ggcagccata tggctagcat gactggtgga cagcaaatgg
      241 gtcgcggatc cgaattctgg agtcatccac aatttgagaa gggcggcggc agtggcggcg
      301 gcagtggcgg cagtgcctgg agccacccgc agttcgaaaa aggcagtatg agcaaaggag
      361 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
      421 acaaattttc tgtccgtgga gagggtgaag gtgatgctac aaacggaaaa ctcaccctta
      481 aatttatttg cactactgga aaactacctg ttccgtggcc aacacttgtc actactctga
      541 cctatggtgt tcaatgcttt tcccgttatc cggatcacat gaaacggcat gactttttca
      601 agagtgccat gcccgaaggt tatgtacagg aacgcactat atctttcaaa gatgacggga
      661 cctacaagac gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt
      721 taaagggtat tgattttaaa gaagatggaa acattcttgg acacaaactc gaatacaact
      781 ttaactcaca caatgtatac atcacggcag acaaacaaaa gaatggaatc aaagctaact
      841 tcaaaattcg ccacaacgtt gaagatggtt ccgttcaact agcagaccat tatcaac
//
