LOCUS       sequence-136433-         940 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for Citrine purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 940)
  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 940)
  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-Citrine (525)
FEATURES             Location/Qualifiers
     source          1..940
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             2..24
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             44..61
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             68..784
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSRLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             819..836
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(889..907)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 ttttgtttaa ctttaagaag gagatataca tatgagagga tcgcatcacc atcaccatca
       61 cggatccatg tctaaaggtg aagaattatt cactggtgtt gtcccaattt tggttgaatt
      121 agatggtgat gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgatgctac
      181 ttacggtaaa ttgaccttaa aatttatttg tactactggt aaattgccag ttccatggcc
      241 aaccttagtc actactttag gttatggttt gatgtgtttt gctagatacc cagatcatat
      301 gaaacaacat gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat
      361 ttttttcaaa gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac
      421 cttagttaat agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg
      481 tcacaaattg gaatacaact ataactctca caatgtttac atcatggctg acaaacaaaa
      541 gaatggtatc aaagttaact tcaaaattag acacaacatt gaagatggtt ctgttcaatt
      601 agctgaccat tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa
      661 ccattactta tcctatcaat ctagattatc caaagatcca aacgaaaaga gagatcacat
      721 ggtcttgtta gaatttgtta ctgctgctgg tattacccat ggtatggatg aattgtacaa
      781 atgagagctc cgtcgacaag cttgcggccg cactcgagca ccaccaccac caccactgag
      841 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      901 aactagcata accccttggg gcctctaaac gggtcttgag
//
