LOCUS       sequence-109752-         959 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 959)
  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 959)
  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..959
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     69..88
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        69..87
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    88..112
                     /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             127..149
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             168..185
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             195..212
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             555..572
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(625..643)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      639..686
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(810..829)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
       61 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
      121 aataattttg tttaacttta agaaggagat ataccatggg cagcagccat catcatcatc
      181 atcacagcag cggcctggtg ccgcgcggca gccatatgtt aatattagat ggggacctac
      241 taaaggacaa gttaaagtta ccggtcattg acaatctgtt tggtaaagaa ctccttgaca
      301 agtttcaaga tgatattaag gacaaatatg gggtggacac caaagacctt aagatcctca
      361 agacatcaga agataaaagg ttttactatg tgtcggtcga tgctggagat ggcgagaaat
      421 gtaaattcaa gattagaaaa gatgtcgatg ttccaaagat ggtcggacgc aaatgtagga
      481 aggacgatga tgatgatgat ggatattagg atccgaattc gagctccgtc gacaagcttg
      541 cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
      601 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
      661 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
      721 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
      781 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
      841 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
      901 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtg
//
