LOCUS       sequence-64525-e        2734 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 2734)
  AUTHORS   Hocine S, Raymond P, Zenklusen D, Chao JA, Singer RH
  TITLE     Single-molecule analysis of gene expression using two-color RNA 
            labeling in live yeast.
  JOURNAL   Nat Methods. 2013 Feb;10(2):119-21. doi: 10.1038/nmeth.2305. Epub 
            2012 Dec 23.
  PUBMED    23263691
REFERENCE   2  (bases 1 to 2734)
  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: 
            "10.1038/nmeth.2305"; journalName: "Nat Methods"; date: "2013-02"; 
            volume: "10"; issue: "2"; pages: "119-21"
COMMENT     Sequence Label: pDZ274 (pLEU MET25pro MCP-2x-yeGFP)
FEATURES             Location/Qualifiers
     source          1..2734
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     33..53
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        35..53
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        104..453
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             489..875
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2
                     /note="binds to a specific stem-loop structure in the viral
                     RNA (Peabody, 1993)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQNRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             927..1640
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             927..1640
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1692..2408
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1692..2405
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2415..2432
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2466..2713
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2482..2500)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
ORIGIN
        1 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
       61 agctggagct ccggatgcaa gggttcgaat cccttagctc tcattatttt ttgctttttc
      121 tcttgaggtc acatgatcgc aaaatggcaa atggcacgtg aagctgtcga tattggggaa
      181 ctgtggtggt tggcaaatga ctaattaagt tagtcaaggc gccatcctca tgaaaactgt
      241 gtaacataat aaccgaagtg tcgaaaaggt ggcaccttgt ccaattgaac acgctcgatg
      301 aaaaaaataa gatatatata aggttaagta aagcgtctgt tagaaaggaa gtttttcctt
      361 tttcttgctc tcttgtcttt tcatctacta tttccttcgt gtaatacagg gtcgtcagat
      421 acatagatac aattctatta cccccatcca tactctagat agagccctca accggagttt
      481 gaagcatggc ttctaacttt actcagttcg ttctcgtcga caatggcgga actggcgacg
      541 tgactgtcgc cccaagcaac ttcgctaacg gggtcgctga atggatcagc tctaactcgc
      601 gttcacaggc ttacaaagta acctgtagcg ttcgtcagag ctctgcgcag aatcgcaaat
      661 acaccatcaa agtcgaggtg cctaaagtgg caacccagac tgttggtggt gtagagcttc
      721 ctgtagccgc atggcgttcg tacctaaata tggaactaac cattccaatt ttcgccacga
      781 attccgactg cgagcttatt gttaaggcaa tgcaaggtct cctaaaagat ggaaacccga
      841 ttccctcagc aatcgcagca aactccggca tctacggatc ccccgggctg caggaattcg
      901 atatcaagct tatcgatacc gtcgacatgt ctaaaggtga agaattattc actggtgttg
      961 tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct gtctccggtg
     1021 aaggtgaagg tgatgctact tacggtaaat tgaccttaaa atttatttgt actactggta
     1081 aattgccagt tccatggcca accttagtca ctactttaac ttatggtgtt caatgttttt
     1141 ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg ccagaaggtt
     1201 atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc agagctgaag
     1261 tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt gattttaaag
     1321 aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac aatgtttaca
     1381 tcatggctga caaacaaaag aatggtatca aagttaactt caaaattaga cacaacattg
     1441 aagatggttc tgttcaatta gctgaccatt atcaacaaaa tactccaatt ggtgatggtc
     1501 cagtcttgtt accagacaac cattacttat ccactcaatc tgccttatcc aaagatccaa
     1561 acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt attacccatg
     1621 gtatggatga attgtacaaa agatcccccg ggctgcagga attcgatatc aagcttatcg
     1681 ataccgtcga catgtctaaa ggtgaagaat tattcactgg tgttgtccca attttggttg
     1741 aattagatgg tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt gaaggtgatg
     1801 ctacttacgg taaattgacc ttaaaattta tttgtactac tggtaaattg ccagttccat
     1861 ggccaacctt agtcactact ttaacttatg gtgttcaatg tttttctaga tacccagatc
     1921 atatgaaaca acatgacttt ttcaagtctg ccatgccaga aggttatgtt caagaaagaa
     1981 ctattttttt caaagatgac ggtaactaca agaccagagc tgaagtcaag tttgaaggtg
     2041 ataccttagt taatagaatc gaattaaaag gtattgattt taaagaagat ggtaacattt
     2101 taggtcacaa attggaatac aactataact ctcacaatgt ttacatcatg gctgacaaac
     2161 aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggttctgttc
     2221 aattagctga ccattatcaa caaaatactc caattggtga tggtccagtc ttgttaccag
     2281 acaaccatta cttatccact caatctgcct tatccaaaga tccaaacgaa aagagagacc
     2341 acatggtctt gttagaattt gttactgctg ctggtattac ccatggtatg gatgaattgt
     2401 acaaataaac cggtcatcat caccatcacc attgagttta aacccgctga tcctagaggg
     2461 ccgcatcatg taattagtta tgtcacgctt acattcacgc cctcccccca catccgctct
     2521 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt
     2581 tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg
     2641 cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgaa
     2701 ggctttaatt tgcaagctgc ggccctgcat tggc
//
