LOCUS       sequence-104794-        1095 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  bacterial expression of N-terminal fusion protein of CNA35 with 
            mAmetrine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1095)
  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 1095)
  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-mAmetrine-CNA35
FEATURES             Location/Qualifiers
     source          1..1095
                     /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"
     primer_bind     complement(280..301)
                     /label=EGFP-N
                     /note="EGFP, reverse 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 cgtgcgcggc gagggcgagg gcgatgccac caacggcaag
      361 ctgaccctga agttcatctg cacctccggc aagctgcccg tgccctggcc caccctcgtg
      421 accaccctgt cttacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
      481 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat ctccttcaag
      541 gacgacggca gctacaggac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      601 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
      661 gagtacaaca tgaacgtgtg ggacgcgtat atcacggccg acaagcagaa gaacggcatc
      721 aaagcgaact tcaagatcga gcacaacgtc gaggacggcg gcgtgcagct cgccgacgcg
      781 taccagcaga acacccccat cggcgacggc tccgtgctgc tgcctgacaa ccactacctg
      841 agcttccaga gcaagctgtt caaagacccc aacgagcagc gcgatcacat ggtcctgctg
      901 gagttcgtta ccgccgccgg gatcactctc gaattccacg gatccgcacg agatatttca
      961 tcaacgaatg ttacagattt aactgtatca ccgtctaaga tagaagatgg tggtaaaacg
     1021 acagtaaaaa tgacgttcga cgataaaaat ggaaaaatac aaaatggtga catgattaaa
     1081 gtggcatggc cgaca
//
