LOCUS       sequence-96374-e        8205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase (Promega) yeast codon-optimized yellow 
            fluorescent protein (yEVenus) fusion driven by RNR1 promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8205)
  AUTHORS   Mazo-Vargas A, Park H, Aydin M, Buchler NE
  TITLE     Measuring fast gene dynamics in single cells with time-lapse 
            luminescence microscopy
  JOURNAL   Mol Biol Cell. 2014 Sep 17. pii: mbc.E14-07-1187.
  PUBMED    25232010
REFERENCE   2  (bases 1 to 8205)
  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: "Mol Biol 
            Cell. 2014 Sep 17. pii: mbc.E14-07-1187."
COMMENT     Sequence Label: pNB788
FEATURES             Location/Qualifiers
     source          1..8205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2241..2261
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2243..2261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             3254..4903
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDVKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3320..3338)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             4916..5632
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDESRE"
     terminator      5653..5840
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5708..5732)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(6210..6229)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6211..6229)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6239..6256)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6239..6255)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6248..6270)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6400..6855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6487..6506)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6697..6718
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6986..7789)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(7790..8010)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(8036..8055)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8155..8177
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctcc
     2281 accgcggtgg cggccgtcac tgtgtatagt agatctgcag ctcagtcaca tgagacatta
     2341 acgtatataa atatataaat atatatatat ccattaaata acgaagtagg acatggtcta
     2401 aaaatagtgc ttctcatgta ttgattttga gtagccataa gaataattat tgatttcaag
     2461 atctttgatc tccttacatc caaaattata ataccacttt atggtatcac gacatcacca
     2521 caacagagaa ggattgccgc atttggatat cgtaaacaaa ggcgttacca tagaaatgta
     2581 ctgattggca gaattactct tcaggagaat ctttcataca aaggaattcc attggggaaa
     2641 atctcgttac caagtcaatg ctgaactttc tatggccttt gtttactatc gttaattatt
     2701 ttacgaccac ttctgggtag aaatatttcg tagccctgga acgagcttgt ttacgcgttt
     2761 tatcccatta tatggcaccc aaatcaaatt taaaaagaaa aaacgcgtaa acagtgtcgg
     2821 gtaagttcat cctctgttac tttaattgct tctttttttg aaattctaag taaacgcgtc
     2881 attttgatcc tcaggacaca gaaatccttg cagaatctta ttgggtgttg aatagaggac
     2941 gcgtaaaaac gatatggaaa tttttttcat atagtgtaga aagaataggt tggcgtaggt
     3001 agtttcgtgt ttgatagaaa cctccaacaa agtctgcagc tcacgtttta gaataacaag
     3061 tttagagttt atcttgttgc ctttgttaag tcagtaccat tgaataaaaa ttatataaag
     3121 gagctaatat ttcattgttg gaaaattact ctaccataat tgaagcatat ctcatccttt
     3181 tcatcctttt caacgcaaga gagacaccaa cgaacaacac tttatttgtt gatatattaa
     3241 catcttaatt aacatggaag acgtcaaaaa cataaagaaa ggcccggcgc cattctatcc
     3301 gctggaagat ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt
     3361 tcctggaaca attgctttta cagatgcaca tatcgaggtg gacatcactt acgctgagta
     3421 cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca
     3481 cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt
     3541 atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa
     3601 cagtatgggc atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat
     3661 tttgaacgtg caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg attctaaaac
     3721 ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt
     3781 taatgaatac gattttgtgc cagagtcctt cgatagggac aagacaattg cactgatcat
     3841 gaactcctct ggatctactg gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg
     3901 cgtgagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc
     3961 gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt
     4021 gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttctgaggag
     4081 ccttcaggat tacaagattc aaagtgcgct gctggtgcca accctattct ccttcttcgc
     4141 caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg cttctggtgg
     4201 cgctcccctc tctaaggaag tcggggaagc ggttgccaag aggttccatc tgccaggtat
     4261 caggcaagga tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga
     4321 tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct
     4381 ggataccggg aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat
     4441 gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg
     4501 atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct tcatcgttga
     4561 ccgcctgaag tctctgatta agtacaaagg ctatcaggtg gctcccgctg aattggaatc
     4621 catcttgctc caacacccca acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga
     4681 cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa
     4741 agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt
     4801 tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag
     4861 agagatcctc ataaaggcca agaagggcgg aaagatcgcc gtgggtgggg gatcaatgtc
     4921 taaaggtgaa gaattattca ctggtgttgt cccaattttg gttgaattag atggtgatgt
     4981 taatggtcac aaattttctg tctccggtga aggtgaaggt gatgctactt acggtaaatt
     5041 gaccttaaaa ttgatttgta ctactggtaa attgccagtt ccatggccaa ccttagtcac
     5101 tactttaggt tatggtttgc aatgttttgc tagataccca gatcatatga aacaacatga
     5161 ctttttcaag tctgccatgc cagaaggtta tgttcaagaa agaactattt ttttcaaaga
     5221 tgacggtaac tacaagacca gagctgaagt caagtttgaa ggtgatacct tagttaatag
     5281 aatcgaatta aaaggtattg attttaaaga agatggtaac attttaggtc acaaattgga
     5341 atacaactat aactctcaca atgtttacat cactgctgac aaacaaaaga atggtatcaa
     5401 agctaacttc aaaattagac acaacattga agatggtggt gttcaattag ctgaccatta
     5461 tcaacaaaat actccaattg gtgatggtcc agtcttgtta ccagacaacc attacttatc
     5521 ctatcaatct gccttatcca aagatccaaa cgaaaagaga gaccacatgg tcttgttaga
     5581 atttgttact gctgctggta ttacccatgg tattgatgaa tctcgagaat aaggcgcgcc
     5641 acttctaaat aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa
     5701 aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc
     5761 ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagtatga ggtcgctctt
     5821 attgaccaca cctctaccgg cagatccgct agggataaca gggtaatata gatctgttta
     5881 gcttgcctcg tccccgccgg gtcacccggc cagcgacatg gaggcccaga ataccctcct
     5941 tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt acatttagcc
     6001 catacatccc catgtataat catttgcatc catacatttt gatggccgca cggcgcgaag
     6061 caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc tcacagacgc
     6121 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag
     6181 gctttaattt gcggccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     6241 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     6301 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     6361 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat
     6421 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     6481 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     6541 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     6601 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     6661 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     6721 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     6781 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     6841 taacgtttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     6901 ccgcataggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac
     6961 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt
     7021 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata
     7081 gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat
     7141 actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc
     7201 aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat
     7261 ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc
     7321 ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg
     7381 ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt
     7441 ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa
     7501 tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta
     7561 gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta
     7621 aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat
     7681 ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa
     7741 caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc
     7801 gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt
     7861 cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt
     7921 ccttctgttc ggagattacc gaatcaaaaa aatttcaaag aaaccgaaat caaaaaaaag
     7981 aataaaaaaa aaatgatgaa ttgaattgaa aagctgtggt atggtgcact ctcagtacaa
     8041 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     8101 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     8161 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcga
//
