LOCUS       sequence-146534-         923 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 923)
  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 923)
  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 I136A, I139A, M142A
FEATURES             Location/Qualifiers
     source          1..923
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     68..87
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        68..86
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    87..111
                     /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             126..148
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             166..183
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             193..210
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             214..246
                     /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(262..285,286..321,322..345)
                     /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(382..410)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
       61 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
      121 ataattttgt ttaactttaa gaaggagata tacatgggca gcagccatca tcatcatcat
      181 cacagcagcg gcctggtgcc gcgcggcagc catatggcta gcatgactgg tggacagcaa
      241 atgggtcgcg gatccgaatt ctggagtcat ccacaatttg agaagggcgg cggcagtggc
      301 ggcggcagtg gcggcagtgc ctggagccac ccgcagttcg aaaaaggcag tatgagcaaa
      361 ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat
      421 gggcacaaat tttctgtccg tggagagggt gaaggtgatg ctacaaacgg aaaactcacc
      481 cttaaattta tttgcactac tggaaaacta cctgttccgt ggccaacact tgtcactact
      541 ctgacctatg gtgttcaatg cttttcccgt tatccggatc acatgaaacg gcatgacttt
      601 ttcaagagtg ccatgcccga aggttatgta caggaacgca ctatatcttt caaagatgac
      661 gggacctaca agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt taatcgtatc
      721 gagttaaagg gtattgattt taaagaagat ggaaacattc ttggacacaa actcgaatac
      781 aactttaact cacacaatgt atacatcacg gcagacaaac aaaagaatgg aatcaaagct
      841 aacttcaaaa ttcgccacaa cgttgaagat ggttccgttc aactagcaga ccattatcaa
      901 caaaatactc caattggcga tgg
//
