LOCUS       sequence-126640-        3330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for building 3-color reporter plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3330)
  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 3330)
  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: pPhlF-BCD20-Citrine-ssrA P64U23C88T18 v1-1 (586)
FEATURES             Location/Qualifiers
     source          1..3330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             158..871
                     /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"
     primer_bind     complement(1103..1120)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1274..1862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1354..1373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2033..2824,2825..2893))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHATRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2656..2675
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2894..2998)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3066..3084
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcattgctgt tcgattcgtt accaattgac atgatacgaa acgtaccgta tcgttaaggt
       61 tactagagaa gcgggcccaa gttcacttaa aaaggagatc aacaatgaaa gcaattttcg
      121 tactgaaaca tcttaatcat gctgaggaaa gtttctaatg tctaaaggtg aagaattatt
      181 cactggtgtt gtcccaattt tggttgaatt agatggtgat gttaatggtc acaaattttc
      241 tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa ttgaccttaa aatttatttg
      301 tactactggt aaattgccag ttccatggcc aaccttagtc actactttag gttatggttt
      361 gatgtgtttt gctagatacc cagatcatat gaaacaacat gactttttca agtctgccat
      421 gccagaaggt tatgttcaag aaagaactat ttttttcaaa gatgacggta actacaagac
      481 cagagctgaa gtcaagtttg aaggtgatac cttagttaat agaatcgaat taaaaggtat
      541 tgattttaaa gaagatggta acattttagg tcacaaattg gaatacaact ataactctca
      601 caatgtttac atcatggctg acaaacaaaa gaatggtatc aaagttaact tcaaaattag
      661 acacaacatt gaagatggtt ctgttcaatt agctgaccat tatcaacaaa atactccaat
      721 tggtgatggt ccagtcttgt taccagacaa ccattactta tcctatcaat ctagattatc
      781 caaagatcca aacgaaaaga gagatcacat ggtcttgtta gaatttgtta ctgctgctgg
      841 tattacccat ggtatggatg aattgtacaa agcagcaaac gacgaaaact acgctttagc
      901 agcttgatga actctcggta ccaaattcca gaaaagaggc ctcccgaaag gggggccttt
      961 tttcgttttg gtccccttga gagtcgggca ttgtcttcgc tccttccggt gggcgcgggg
     1021 catgactatc gtcgccgcac ttatgactgt gttctttatc atgcaactcg taggacaggt
     1081 gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1141 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1201 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1261 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1321 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1381 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1441 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1501 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1561 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1621 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1681 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1741 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1801 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1861 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1921 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1981 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2041 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2101 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2161 caatgatacc gcgtgaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2221 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2281 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2341 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2401 gttcccaacg atcaaggcga gttgcatgat cccccatgtt gtgcaaaaaa gcggttagct
     2461 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2521 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2581 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2641 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     2701 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     2761 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     2821 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     2881 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2941 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3001 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     3061 ataaaaatag gcgtatcacg aggccctttc gtgttcaaga attctggcga atcctctgac
     3121 cagccagaaa acgacctttc tgtggtgaaa ccggatgctg caattcagag cgccagcaag
     3181 tgggggacag cagatgacct gaccgccgca gagtggatgt ttgacatggt gatgactatc
     3241 gcaccatcag ccagaaaacc gaattttgct gggtgggcta acgatatccg cctgatgcgt
     3301 gaacgtgacg gacgtaacga agacaaccac
//
