LOCUS       sequence-146513-         863 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses a triple mutant truncation of Hook3 in bacteria for 
            purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 863)
  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 863)
  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 Q147A, M151A, I154A
FEATURES             Location/Qualifiers
     source          1..863
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     69..88
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        69..87
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    88..112
                     /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             127..149
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             167..184
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             194..211
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             215..247
                     /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(263..286,287..322,323..346)
                     /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(383..411)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
       61 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
      121 aataattttg tttaacttta agaaggagat atacatgggc agcagccatc atcatcatca
      181 tcacagcagc ggcctggtgc cgcgcggcag ccatatggct agcatgactg gtggacagca
      241 aatgggtcgc ggatccgaat tctggagtca tccacaattt gagaagggcg gcggcagtgg
      301 cggcggcagt ggcggcagtg cctggagcca cccgcagttc gaaaaaggca gtatgagcaa
      361 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
      421 tgggcacaaa ttttctgtcc gtggagaggg tgaaggtgat gctacaaacg gaaaactcac
      481 ccttaaattt atttgcacta ctggaaaact acctgttccg tggccaacac ttgtcactac
      541 tctgacctat ggtgttcaat gcttttcccg ttatccggat cacatgaaac ggcatgactt
      601 tttcaagagt gccatgcccg aaggttatgt acaggaacgc actatatctt tcaaagatga
      661 cgggacctac aagacgcgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat
      721 cgagttaaag ggtattgatt ttaaagaaga tggaaacatt cttggacaca aactcgaata
      781 caactttaac tcacacaatg tatacatcac ggcagacaaa caaaagaatg gaatcaaagc
      841 taacttcaaa attcgccaca acg
//
