LOCUS       sequence-59440-e         977 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 977)
  AUTHORS   Phong C, Markson JS, Wilhoite CM, Rust MJ
  TITLE     Robust and tunable circadian rhythms from differentially sensitive 
            catalytic domains.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Jan 15;110(3):1124-9. doi: 
            10.1073/pnas.1212113110. Epub 2012 Dec 31.
  PUBMED    23277568
REFERENCE   2  (bases 1 to 977)
  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.1212113110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-01-15- 15"; volume: "110"; issue: "3"; pages: "1124-9"
COMMENT     Sequence Label: KaiB
FEATURES             Location/Qualifiers
     source          1..977
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(63..107)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(108..125)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(477..500)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(510..527)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             541..563
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    578..602
                     /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(602..621)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(603..621)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(709..728)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cttcctttcg ggctttgttt agcagcctag gttaattaag cctcgagagc agcagaagta
       61 gagctgtcca tgtgctggcg ttcgaattta gcagcagcgg tttctttact accgcgtggc
      121 accagagcga gctctgcggc cgcaagcttc tattagaagt cgtcggaatc ttgaagttcg
      181 ccgtagagta aatcaaggcc aatcaaaact ttctcacggt cggataaatc accaataatc
      241 cgtcggacag gcagtggtag aaccttggcg agggttggcg tcgctaggat tttatcctct
      301 tctgccaact gaggattttt gagaacatcg atcaccttta gagcataaac accttgaaat
      361 tcaacttcga gaatgttctt gagcgttttg agggcacgga ctgagtttgg agtattgccg
      421 gcgacgtaga gcttgagaat gtaggtttta cgagggctca tcggatcctg gtacccgggt
      481 ccctgaaaga ggacttcaag agccgcggag tgatggtggt ggtgatgtgc catatgtata
      541 tctccttctt aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt
      601 cccctatagt gagtcgtatt aatttcgcgg gatcgagatc gatctcgatc ctctacgccg
      661 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
      721 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg
      781 tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac
      841 cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc
      901 aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg caaaaccttt
      961 cgcggtatgg catgata
//
