LOCUS       sequence-136431-         850 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for Cerulean purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 850)
  AUTHORS   Niederholtmeyer H, Sun ZZ, Hori Y, Yeung E, Verpoorte A, Murray RM, 
            Maerkl SJ
  TITLE     Rapid cell-free forward engineering of novel genetic ring 
            oscillators.
  JOURNAL   Elife. 2015 Oct 2;4. doi: 10.7554/eLife.09771.
  PUBMED    26430766
REFERENCE   2  (bases 1 to 850)
  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: "Elife."; 
            date: "2015-10-2"; volume: "4. doi"; pages: " 10.7554/eLife.09771"
COMMENT     Sequence Label: pET21a(+)-Histag-Cerulean (526)
FEATURES             Location/Qualifiers
     source          1..850
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(27..44)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(82..795)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(82..795)
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(122..142)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     737..765
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     CDS             complement(802..819)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 cgggctttgt tagcagccgg atctcagtgg tggtggtggt ggtgctcgag tgcggccgca
       61 agcttgtcga cggagctctt atttgtatag ttcatccatg cctagtgtaa tcccagcagc
      121 tgttacaaac tcaagaagga ccatgtgatc tctcttttcg ttgggatctt tcgaaagggc
      181 agattgtgtg gacaggtaat ggttgtctgg taaaaggaca gggccatcgc caattggagt
      241 attttgttga taatggtctg ctagttgaac gcttccatct tcaatgttgt gtctaatttt
      301 gaaattagct ttgattccat tcttttgttt gtctgcagtg atgtatacat tatctgaaat
      361 agcgttgtat tccaatttgt gtccaagaat gtttccatct tctttaaaat caataccttt
      421 taactcgatt ctattaacaa gggtatcacc ttcaaacttg acttcagcac gtgtcttgta
      481 gttcccgtca tctttgaaaa atatagttct ttcctgtaca taaccttcgg gcatggcact
      541 cttgaaaaag tcatgctgtt tcatatgatc tgggtatcta gcaaagcatt gaacacccca
      601 agtcaaagta gtgacaagtg ttggccatgg aacaggtagt tttccagtag tgcaaataaa
      661 tttaagggta agttttccgt atgttgcatc accttcaccc tctccactga cagaaaattt
      721 gtgcccatta acatcaccat ctaattcaac aagaattggg acaactccag tgaaaagttc
      781 ttctccttta ctcatggatc cgtgatggtg atggtgatgc gatcctctca tatgtatatc
      841 tccttcttaa
//
