LOCUS       sequence-96373-e        8053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase (Promega) yeast codon-optimized yellow 
            fluorescent protein (yEVenus) fusion with PEST degron driven by SIC1
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8053)
  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 8053)
  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: pNB785
FEATURES             Location/Qualifiers
     source          1..8053
                     /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"
     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             2568..4217
                     /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(2634..2652)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      5501..5688
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5556..5580)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(6058..6077)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6059..6077)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6087..6104)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6087..6103)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6096..6118)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6248..6703
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6335..6354)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6545..6566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6834..7637)
                     /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(7638..7858)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7884..7903)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8003..8025
                     /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 tgtgagttac ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcctat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctcc
     2281 tccggcgatc aaacatacca ttcccccctc aaatcaatat gattaagtca cgataatatc
     2341 tgcactattg ccgtgtttcc ttctccatcg acagcgtcga tcctgtctat tatagccagc
     2401 acactgaaaa tttcaatggc tggcttgttg ttcgcgtcgc aacttttgat atttataaaa
     2461 agagagaagg catatccaaa ggataaggta ttgtctttgt aaatcaatca accaaacctc
     2521 tacggaattt tgacccttga agcagggact attacacgtt aattaacatg gaagacgtca
     2581 aaaacataaa gaaaggcccg gcgccattct atccgctgga agatggaacc gctggagagc
     2641 aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct tttacagatg
     2701 cacatatcga ggtggacatc acttacgctg agtacttcga aatgtccgtt cggttggcag
     2761 aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact
     2821 ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca gttgcgcccg
     2881 cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gggcatttcg cagcctaccg
     2941 tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa aagctcccaa
     3001 tcatccaaaa aattattatc atggattcta aaacggatta ccagggattt cagtcgatgt
     3061 acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt gtgccagagt
     3121 ccttcgatag ggacaagaca attgcactga tcatgaactc ctctggatct actggtctgc
     3181 ctaaaggtgt cgctctgcct catagaactg cctgcgtgag attctcgcat gccagagatc
     3241 ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt ccattccatc
     3301 acggttttgg aatgtttact acactcggat atttgatatg tggatttcga gtcgtcttaa
     3361 tgtatagatt tgaagaagag ctgtttctga ggagccttca ggattacaag attcaaagtg
     3421 cgctgctggt gccaacccta ttctccttct tcgccaaaag cactctgatt gacaaatacg
     3481 atttatctaa tttacacgaa attgcttctg gtggcgctcc cctctctaag gaagtcgggg
     3541 aagcggttgc caagaggttc catctgccag gtatcaggca aggatatggg ctcactgaga
     3601 ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg gtcggtaaag
     3661 ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg ctgggcgtta
     3721 atcaaagagg cgaactgtgt gtgagaggtc ctatgattat gtccggttat gtaaacaatc
     3781 cggaagcgac caacgccttg attgacaagg atggatggct acattctgga gacatagctt
     3841 actgggacga agacgaacac ttcttcatcg ttgaccgcct gaagtctctg attaagtaca
     3901 aaggctatca ggtggctccc gctgaattgg aatccatctt gctccaacac cccaacatct
     3961 tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc gccgccgttg
     4021 ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac gtcgccagtc
     4081 aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag
     4141 gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag gccaagaagg
     4201 gcggaaagat cgccgtgggt gggggatcaa tgtctaaagg tgaagaatta ttcactggtg
     4261 ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt tctgtctccg
     4321 gtgaaggtga aggtgatgct acttacggta aattgacctt aaaattgatt tgtactactg
     4381 gtaaattgcc agttccatgg ccaaccttag tcactacttt aggttatggt ttgcaatgtt
     4441 ttgctagata cccagatcat atgaaacaac atgacttttt caagtctgcc atgccagaag
     4501 gttatgttca agaaagaact atttttttca aagatgacgg taactacaag accagagctg
     4561 aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt attgatttta
     4621 aagaagatgg taacatttta ggtcacaaat tggaatacaa ctataactct cacaatgttt
     4681 acatcactgc tgacaaacaa aagaatggta tcaaagctaa cttcaaaatt agacacaaca
     4741 ttgaagatgg tggtgttcaa ttagctgacc attatcaaca aaatactcca attggtgatg
     4801 gtccagtctt gttaccagac aaccattact tatcctatca atctgcctta tccaaagatc
     4861 caaacgaaaa gagagaccac atggtcttgt tagaatttgt tactgctgct ggtattaccc
     4921 atggtattga tgaatctcga gaagcatcca acttgaacat ttcgagaaag cttaccatat
     4981 caaccccatc atgctctttc gaaaattcaa atagcacatc cattccttcg cccgcttcct
     5041 catctcaaag ccacactcca atgagaaaca tgagctcact ctctgataac agcgttttca
     5101 gccggaatat ggaacaatca tcaccaatca ctccaagtat gtaccaattt ggtcagcagc
     5161 agtcaaacag tatatgtggt agcaccgtta gtgtgaatag tctggtgaat acaaataaca
     5221 aacaaaggat ctacgaacaa atcacgggtc ctaacagcaa taacgcaacc aatgattata
     5281 ttgatttgct aaacctaaat gagtctaaca aggaaaacca aaatcccgca acggcgcatt
     5341 acctcaatgg gggcccaccc aagacaagct tcattaacca tggaatgttc ccctcgccaa
     5401 ctgggaccat aaatagcggt aaatctagca gtgcctcatc tttaatttct tttggtatgg
     5461 gcaataccca agtaatatag ggcgcgccac ttctaaataa gcgaatttct tatgatttat
     5521 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     5581 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     5641 ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca gatccgctag
     5701 ggataacagg gtaatataga tctgtttagc ttgcctcgtc cccgccgggt cacccggcca
     5761 gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc tcaggggcat
     5821 gatgtgactg tcgcccgtac atttagccca tacatcccca tgtataatca tttgcatcca
     5881 tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg
     5941 agcagggaaa cgctcccctc acagacgcgt gtacgcatgt aacattatac tgaaaacctt
     6001 gcttgagaag gttttgggac gctcgaaggc tttaatttgc ggccggtacc caattcgccc
     6061 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     6121 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     6181 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     6241 tggcgcgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     6301 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     6361 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6421 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     6481 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6541 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6601 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6661 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcctgatg cggtattttc
     6721 tccttacgca tctgtgcggt atttcacacc gcatagggta ataactgata taattaaatt
     6781 gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt
     6841 tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct
     6901 accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct
     6961 gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct
     7021 aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct
     7081 ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc
     7141 ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg
     7201 cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg
     7261 cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca
     7321 gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa
     7381 aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca
     7441 gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac
     7501 tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg
     7561 tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta
     7621 tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg
     7681 gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat
     7741 ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa
     7801 tttcaaagaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaattgaaaa
     7861 gctgtggtat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     7921 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     7981 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     8041 ccgaaacgcg cga
//
