LOCUS       sequence-93663-e         729 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expresses CKK domain of CAMSAP2 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 729)
  AUTHORS   Hendershott MC, Vale RD
  TITLE     Regulation of microtubule minus-end dynamics by CAMSAPs and 
            Patronin.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Apr 22;111(16):5860-5. doi: 
            10.1073/pnas.1404133111. Epub 2014 Mar 26.
  PUBMED    24706919
REFERENCE   2  (bases 1 to 729)
  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: 
            "10.1073/pnas.1404133111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-04-22- 22"; volume: "111"; issue: "16"; pages: "5860-5"
COMMENT     Sequence Label: pET-28a+GFP-CAMSAP2 CKK
FEATURES             Location/Qualifiers
     source          1..729
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(11..34,35..70,71..94))
                     /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"
     CDS             complement(110..142)
                     /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             complement(146..163)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(173..190)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             208..230
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    245..269
                     /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)."
     primer_bind     complement(269..288)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(270..288)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(372..391)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        597..674
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(694..713)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctatactgcc tttttcgaac tgcgggtggc tccaggcact gccgccactg ccgccgccac
       61 tgccgccgcc cttctcaaat tgtggatgac tccagaattc ggatccgcga cccatttgct
      121 gtccaccagt catgctagcc atatggctgc cgcgcggcac caggccgctg ctgtgatgat
      181 gatgatgatg gctgctgccc atgtatatct ccttcttaaa gttaaacaaa attatttcta
      241 gaggggaatt gttatccgct cacaattccc ctatagtgag tcgtattaat ttcgcgggat
      301 cgagatctcg atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
      361 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
      421 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
      481 gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
      541 actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag atcccggaca
      601 ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
      661 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
      721 cttatcaga
//
