LOCUS       sequence-92019-e        2685 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 2685)
  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 2685)
  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..2685
                     /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"
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 cgattacggg cagcccacca accacttcta cctgcatgag
      961 tcatcgccgc agcctcaagc tcatcagcat ccgcagcgca ggacttgggc tcactccgca
     1021 gcagccgccg cttatgaaca acagcaacag atccagcctt ccctggtgga tgtgaatgcc
     1081 tggcagacgc agcagcacca aaagcagaag cagacctgga tgaacaggcc gccctcgagt
     1141 gcgggagccc cgagtcctgg cagctttatg ctgcaccaga acggtggagg aggaggtggt
     1201 ggcggtggtg gtggcgagct acagcacctg tttcaggtac aggcctcgcc acagcatggc
     1261 caacggcagg ttagtggatc caatggcgtg cagcgccagc agtcgttgac caacttgcga
     1321 gacaatcgct cgcccaaggc accacaaaac atgggaatgc ccatgggtat gccaatgcag
     1381 caagaggaca tgatggcacc gcagagtatt tgctttatcg gtgacgagga ggatgttgat
     1441 gagctggagc ggaacattat cgaatctatg cagtcgacgc acatctccga ctttgtacac
     1501 caacagcagc agcagcacca acaccaacag caacttcagc agcaacaacg gttgcagggg
     1561 cacagcggac gaggtagcag ctcggaggat tatgacagcg gggagatgat ctccaacaag
     1621 ctgaacatca ccagcggcaa tctcacctat cgcataccct cgccatcccg tccctccatc
     1681 caagccaaca gcttccagga tccccgagcc atggcagctg cttccggtgg cgaggaccag
     1741 ccgcccgaga agggtttcta catctccttc gacgatgagc agcccaaacg acccaagcca
     1801 cctctgcgcg ccaagcgatc gcctaaaaag gagtctccac caggaagtcg ggacagcgtc
     1861 gataaccagg cgaccctgaa acgtgaatcg ctaagtcatc tgcacaacaa caacaatatt
     1921 ggattcggaa atgatgatgt caacagcaaa ccggtgacca ggcacagtat ccatggccta
     1981 aacaactcca acagtgtcaa atctcccgga aatgccacgt acaacaagta cacggatgag
     2041 ccgcccatcc aactgcgaca gttggccgtt tcgggagcaa tgtcaccaac tagtaacgaa
     2101 cgtcatcact tggacgatgt cagcaatcag tcaccgcagc agacgcagca accaatgtcg
     2161 cccacgcgac tccaacagag cagcaacaac gcagaggcgg ccaagaacaa ggcgctggtc
     2221 atcggagcag attccaccaa tttggatccg gaatctgttg atgagatgga gcggcgcaag
     2281 gagaaaatca tgctgctgtc tttgcaacgt cgccagcaac aggaggaggc gaaggcgcgc
     2341 aaagagattg aggcttccca gaagcgagaa aaggagcgcg aaaaggagga ggaacgagcg
     2401 cggaaaaagg aggagcaaat ggcacggcga gcggccattt tggagcagca cagactcaag
     2461 aaagccattg aagaggccga gcgagagggt aaaaccctgg atcggcccga tctgcacgtg
     2521 aagctgcaat cgcattcatc cacctcaacg accccacgac tgaggcagca gcgcaccacg
     2581 cgtcccaggc ctaagacaat tcacgtggac gatgccagcg tggacatcag cgaggcttca
     2641 agcatctcta gtcggggcaa aaaaggctca agctcgaatc tatag
//
