LOCUS       sequence-136430-         847 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 847)
  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 847)
  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..847
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             5..27
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             47..64
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             71..784
                     /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             71..784
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(101..129)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     724..744
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             822..839
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 taattttgtt taactttaag aaggagatat acatatgaga ggatcgcatc accatcacca
       61 tcacggatcc atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga
      121 attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc
      181 aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg
      241 gccaacactt gtcactactt tgacttgggg tgttcaatgc tttgctagat acccagatca
      301 tatgaaacag catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac
      361 tatatttttc aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga
      421 tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct
      481 tggacacaaa ttggaataca acgctatttc agataatgta tacatcactg cagacaaaca
      541 aaagaatgga atcaaagcta atttcaaaat tagacacaac attgaagatg gaagcgttca
      601 actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga
      661 caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagatca
      721 catggtcctt cttgagtttg taacagctgc tgggattaca ctaggcatgg atgaactata
      781 caaataagag ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac caccaccact
      841 gagatcc
//
