LOCUS       sequence-109754-         879 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  bacterial expression of ranaspumin 2 as a His-tagged fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 879)
  AUTHORS   Mackenzie CD, Smith BO, Meister A, Blume A, Zhao X, Lu JR, Kennedy 
            MW, Cooper A
  TITLE     Ranaspumin-2: structure and function of a surfactant protein from 
            the foam nests of a tropical frog.
  JOURNAL   Biophys J. 2009 Jun 17;96(12):4984-92. doi: 
            10.1016/j.bpj.2009.03.044.
  PUBMED    19527658
REFERENCE   2  (bases 1 to 879)
  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: 
            "10.1016/j.bpj.2009.03"; journalName: "Biophys J"; date: 
            "2009-06-17- 17"; volume: "96"; issue: "12"; pages: "4984-92"
COMMENT     Sequence Label: pET28-Rsn2
FEATURES             Location/Qualifiers
     source          1..879
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(39..56)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(399..416)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(426..443)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             462..484
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    499..523
                     /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(523..542)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(524..542)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(626..645)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tcagcttcct ttcgggcttt gttagcagcc ggatctcagt ggtggtggtg gtggtgctcg
       61 agtgcggccg caagcttgtc gacggagctc gaattcggat cctaatatcc atcatcatca
      121 tcatcgtcct tcctacattt gcgtccgacc atctttggaa catcgacatc ttttctaatc
      181 ttgaatttac atttctcgcc atctccagca tcgaccgaca catagtaaaa ccttttatct
      241 tctgatgtct tgaggatctt aaggtctttg gtgtccaccc catatttgtc cttaatatca
      301 tcttgaaact tgtcaaggag ttctttacca aacagattgt caatgaccgg taactttaac
      361 ttgtccttta gtaggtcccc atctaatatt aacatatggc tgccgcgcgg caccaggccg
      421 ctgctgtgat gatgatgatg atggctgctg cccatggtat atctccttct taaagttaaa
      481 caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat
      541 taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc
      601 ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg
      661 gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg
      721 gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc
      781 aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt
      841 cgagatcccg gacaccatcg aatggcgcaa aacctttcg
//
