LOCUS       sequence-91997-e        1488 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expresses CKK domain of CAMSAP3 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1488)
  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 1488)
  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-CAMSAP3 CKK
FEATURES             Location/Qualifiers
     source          1..1488
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             13..30
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             40..57
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             61..93
                     /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(109..132,133..168,169..192)
                     /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             join(199..201,202..912)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             199..912
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(229..257)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     852..872
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             919..939
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat
       61 atggctagca tgactggtgg acagcaaatg ggtcgcggat ccgaattctg gagtcatcca
      121 caatttgaga agggcggcgg cagtggcggc ggcagtggcg gcagtgcctg gagccacccg
      181 cagttcgaaa aaggcagtat gagcaaagga gaagaacttt tcactggagt tgtcccaatt
      241 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtccgtgg agagggtgaa
      301 ggtgatgcta caaacggaaa actcaccctt aaatttattt gcactactgg aaaactacct
      361 gttccgtggc caacacttgt cactactctg acctatggtg ttcaatgctt ttcccgttat
      421 ccggatcaca tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
      481 gaacgcacta tatctttcaa agatgacggg acctacaaga cgcgtgctga agtcaagttt
      541 gaaggtgata cccttgttaa tcgtatcgag ttaaagggta ttgattttaa agaagatgga
      601 aacattcttg gacacaaact cgagtacaac tttaactcac acaatgtata catcacggca
      661 gacaaacaaa agaatggaat caaagctaac ttcaaaattc gccacaacgt tgaagatggt
      721 tccgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
      781 ttaccagaca accattacct gtcgacacaa tctgtccttt cgaaagatcc caacgaaaag
      841 cgtgaccaca tggtccttct tgagtttgta actgctgctg ggattacaca tggcatggat
      901 gagctctaca aaggatccga aaatctgtat tttcaggggg gtggaggtgg tacacccaat
      961 aaccttggtg tgaagaggcc cacctctcgg gccccttctc catcaggcct catgtctcca
     1021 agccgcctgc ctggcagtcg agaacgtgac tgggagaatg gaagcaatgc atcctcccca
     1081 gcatcagtgc ctgagtacac aggtccccgg ctatacaagg aacccagcgc caagtctaac
     1141 aagtttatca tccacaatgc cttgtcacac tgctgcctgg caggcaaggt gaatgagccc
     1201 cagaagaaca gaattctaga ggaaatcgag aaaagcaagg ccaaccactt cctgattctc
     1261 tttcgggact caagctgcca gttccgggcc ctctacactc tgtctgggga gacagaggag
     1321 ctatcgaggc tggcaggcta tgggcctcgg accgtcactc ctgccatggt cgaaggcatc
     1381 tataagtaca actcggatcg caaacggttc acccagatcc ctgctaaaac catgtctatg
     1441 agtgtagacg ccttcactat ccagggacac ctttggcaaa gcaagtag
//
