LOCUS       sequence-96368-e        7510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast codon-optimized yellow fluorescent protein (yEVenus) with PEST
            degron driven by SIC1 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7510)
  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 7510)
  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: pNB813
FEATURES             Location/Qualifiers
     source          1..7510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(663..1757)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1770..2174)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2474..2929)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2611..2632)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2823..2842
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3051..3072
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3087..3092,3093..3110,3111..3117)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3125..3141
                     /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     3130..3152
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3149..3165
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3149..3165
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3184..3204
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3186..3204
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             3511..4224
                     /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
                     FVTAAGITHGIDELYK"
     terminator      4782..4969
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4837..4861)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5339..5358)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5340..5358)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5368..5385)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5368..5384)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5377..5399)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5520..5537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5691..6279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5771..5790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6450..7241,7242..7310))
                     /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     7073..7092
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7311..7415)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7483..7501
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc
      241 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca
      301 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat
      361 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc
      421 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc
      481 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt
      541 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg
      601 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct
      661 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg
      721 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct
      781 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac
      841 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat
      901 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc
      961 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg
     1021 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca
     1081 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc
     1141 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata
     1201 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact
     1261 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc
     1321 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca
     1381 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt
     1441 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca
     1501 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg
     1561 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca
     1621 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga
     1681 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc
     1741 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata
     1801 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat
     1861 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat
     1921 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct
     1981 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca
     2041 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat
     2101 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga
     2161 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg
     2221 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt
     2281 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg
     2341 atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt ggatatacta
     2401 gaagttctcc tcgaccgtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2461 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     2521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2701 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2761 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2821 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2881 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat
     2941 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     3001 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     3061 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     3121 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     3181 cgcgcaatta accctcacta aagggaacaa aagctggagc tcctccggcg atcaaacata
     3241 ccattccccc ctcaaatcaa tatgattaag tcacgataat atctgcacta ttgccgtgtt
     3301 tccttctcca tcgacagcgt cgatcctgtc tattatagcc agcacactga aaatttcaat
     3361 ggctggcttg ttgttcgcgt cgcaactttt gatatttata aaaagagaga aggcatatcc
     3421 aaaggataag gtattgtctt tgtaaatcaa tcaaccaaac ctctacggaa ttttgaccct
     3481 tgaagcaggg actattacac gttaattaac atgtctaaag gtgaagaatt attcactggt
     3541 gttgtcccaa ttttggttga attagatggt gatgttaatg gtcacaaatt ttctgtctcc
     3601 ggtgaaggtg aaggtgatgc tacttacggt aaattgacct taaaattgat ttgtactact
     3661 ggtaaattgc cagttccatg gccaacctta gtcactactt taggttatgg tttgcaatgt
     3721 tttgctagat acccagatca tatgaaacaa catgactttt tcaagtctgc catgccagaa
     3781 ggttatgttc aagaaagaac tatttttttc aaagatgacg gtaactacaa gaccagagct
     3841 gaagtcaagt ttgaaggtga taccttagtt aatagaatcg aattaaaagg tattgatttt
     3901 aaagaagatg gtaacatttt aggtcacaaa ttggaataca actataactc tcacaatgtt
     3961 tacatcactg ctgacaaaca aaagaatggt atcaaagcta acttcaaaat tagacacaac
     4021 attgaagatg gtggtgttca attagctgac cattatcaac aaaatactcc aattggtgat
     4081 ggtccagtct tgttaccaga caaccattac ttatcctatc aatctgcctt atccaaagat
     4141 ccaaacgaaa agagagacca catggtcttg ttagaatttg ttactgctgc tggtattacc
     4201 catggtattg atgaattgta caaagcatcc aacttgaaca tttcgagaaa gcttaccata
     4261 tcaaccccat catgctcttt cgaaaattca aatagcacat ccattccttc gcccgcttcc
     4321 tcatctcaaa gccacactcc aatgagaaac atgagctcac tctctgataa cagcgttttc
     4381 agccggaata tggaacaatc atcaccaatc actccaagta tgtaccaatt tggtcagcag
     4441 cagtcaaaca gtatatgtgg tagcaccgtt agtgtgaata gtctggtgaa tacaaataac
     4501 aaacaaagga tctacgaaca aatcacgggt cctaacagca ataacgcaac caatgattat
     4561 attgatttgc taaacctaaa tgagtctaac aaggaaaacc aaaatcccgc aacggcgcat
     4621 tacctcaatg ggggcccacc caagacaagc ttcattaacc atggaatgtt cccctcgcca
     4681 actgggacca taaatagcgg taaatctagc agtgcctcat ctttaatttc ttttggtatg
     4741 ggcaataccc aagtaatata gggcgcgcca cttctaaata agcgaatttc ttatgattta
     4801 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact
     4861 cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc
     4921 tttctcaggt atagtatgag gtcgctctta ttgaccacac ctctaccggc agatccgcta
     4981 gggataacag ggtaatatag atctgtttag cttgcctcgt ccccgccggg tcacccggcc
     5041 agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca
     5101 tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc
     5161 atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc
     5221 gagcagggaa acgctcccct cacagacgcg tgtacgcatg taacattata ctgaaaacct
     5281 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccggtac ccaattcgcc
     5341 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     5401 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctgcat
     5461 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5521 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5581 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5641 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5701 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5761 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5821 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5881 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5941 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6001 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6061 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6121 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6181 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6241 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6301 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6361 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6421 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6481 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6541 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6601 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6661 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6721 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6781 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6841 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6901 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6961 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7021 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7081 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7141 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7201 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7261 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7321 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7381 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7441 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     7501 ccctttcgtc
//
