LOCUS       sequence-104760-        1050 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  bacterial expression of N-terminal fusion protein of CNA35 with 
            m-Turquoise-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1050)
  AUTHORS   Aper SJ, van Spreeuwel AC, van Turnhout MC, van der Linden AJ, 
            Pieters PA, van der Zon NL, de la Rambelje SL, Bouten CV, Merkx M
  TITLE     Colorful Protein-Based Fluorescent Probes for Collagen Imaging.
  JOURNAL   PLoS One. 2014 Dec 9;9(12):e114983. doi: 
            10.1371/journal.pone.0114983. eCollection 2014.
  PUBMED    25490719
REFERENCE   2  (bases 1 to 1050)
  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"; journalName: "PLoS One.";
            date: "2014-12-9"; volume: "9(12):e114983. doi"; pages: " 
            10.1371/journal.pone.0114983. eCollection 2014"
COMMENT     Sequence Label: pET28a-mTurquoise2-CNA35
FEATURES             Location/Qualifiers
     source          1..1050
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     67..86
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        67..85
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    86..110
                     /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             125..147
                     /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"
     regulatory      229..238
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(235..237,238..240,241..951)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(280..301)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(541..560)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     888..909
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
       61 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
      121 taattttgtt taactttaag aaggagatat accatgggca gcagccatca tcatcatcat
      181 cacagcagcg gcctggtgcc gcgcggcagc catatggcta gcccggtcgc caccatggtg
      241 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      301 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      361 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      421 accaccctgt cctggggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
      481 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      541 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      601 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
      661 gagtacaact actttagcga caacgtctat atcaccgccg acaagcagaa gaacggcatc
      721 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
      781 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
      841 agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
      901 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa ggaattccac
      961 ggatccgcac gagatatttc atcaacgaat gttacagatt taactgtatc accgtctaag
     1021 atagaagatg gtggtaaaac gacagtaaaa
//
