LOCUS       sequence-59441-e         982 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 982)
  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 982)
  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-FLAG
FEATURES             Location/Qualifiers
     source          1..982
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(45..68)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(375..398)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(join(408..422,423..431))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(435..467)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(471..488)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             508..530
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(561..580)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(562..580)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(665..682)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(916..935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcgggct ttgtttagca gccggatcaa gcttcgaatt cttatttatc atcatcatct
       61 ttataatcga agtcgtcgga atcttgaagt tcgccgtaga gtaaatcaag gccaatcaaa
      121 actttctcac ggtcggataa atcaccaata atccgtcgga caggcagtgg tagaaccttg
      181 gcgagggttg gcgtcgctag gattttatcc tcttctgcca actgaggatt tttgagaaca
      241 tcgatcacct ttagagcata aacaccttga aattcaactt cgagaatgtt cttgagcgtt
      301 ttgagggcac ggactgagtt tggagtattg ccggcgacgt agagcttgag aatgtaggtt
      361 ttacgagggc tcatgggccc ctggaacaga acttccagtc ccggatcctt atcgtcatcg
      421 tcgtacagat cccgacccat ttgctgtcca ccagtcatgc tagccatacc atgatgatga
      481 tgatgatgag aaccccgcat atgtatatct ccttcttaaa gttaaacaaa attatttcta
      541 gagggaaacc gttgtggtct ccctatagtg agtcgtatta atttcgcggg atcgagatcc
      601 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
      661 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      721 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      781 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      841 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      901 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      961 ctgttccgac cctgccgctt ac
//
