LOCUS       sequence-93923-e         750 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  for E. coli expression of Patronin CC truncation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 750)
  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 750)
  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-Patronin CC 868–1457
FEATURES             Location/Qualifiers
     source          1..750
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(3..26,27..62,63..86))
                     /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(102..134)
                     /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(138..155)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(165..182)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             200..222
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    237..261
                     /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(261..280)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(262..280)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(364..383)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        589..666
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cctttttcga actgcgggtg gctccaggca ctgccgccac tgccgccgcc actgccgccg
       61 cccttctcaa attgtggatg actccagaat tcggatccgc gacccatttg ctgtccacca
      121 gtcatgctag ccatatggct gccgcgcggc accaggccgc tgctgtgatg atgatgatga
      181 tggctgctgc ccatgtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
      241 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatct
      301 cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg
      361 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga
      421 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca
      481 tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg
      541 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa
      601 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
      661 gtgaatgtga aaccagtaac gttatacgat gtcgcaggag tatgccggtg tctcttatca
      721 gaccgtttcc cgcgtggtga accaggccag
//
