LOCUS       sequence-96366-e        7939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast codon-optimized yellow fluorescent protein (yEVenus) with PEST
            degron driven by RNR1 promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7939)
  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 7939)
  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: pNB812
FEATURES             Location/Qualifiers
     source          1..7939
                     /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             3254..3967
                     /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      4525..4712
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4580..4604)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4780..4796)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4781..4797)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4821..4840)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4822..4840)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4850..4867)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4850..4866)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4859..4881)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5011..5466
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5098..5117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5308..5329
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5766..6170
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6183..7277
                     /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"
     primer_bind     complement(7770..7789)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7889..7911
                     /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 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 aacatgtcta aaggtgaaga attattcact ggtgttgtcc caattttggt
     3301 tgaattagat ggtgatgtta atggtcacaa attttctgtc tccggtgaag gtgaaggtga
     3361 tgctacttac ggtaaattga ccttaaaatt gatttgtact actggtaaat tgccagttcc
     3421 atggccaacc ttagtcacta ctttaggtta tggtttgcaa tgttttgcta gatacccaga
     3481 tcatatgaaa caacatgact ttttcaagtc tgccatgcca gaaggttatg ttcaagaaag
     3541 aactattttt ttcaaagatg acggtaacta caagaccaga gctgaagtca agtttgaagg
     3601 tgatacctta gttaatagaa tcgaattaaa aggtattgat tttaaagaag atggtaacat
     3661 tttaggtcac aaattggaat acaactataa ctctcacaat gtttacatca ctgctgacaa
     3721 acaaaagaat ggtatcaaag ctaacttcaa aattagacac aacattgaag atggtggtgt
     3781 tcaattagct gaccattatc aacaaaatac tccaattggt gatggtccag tcttgttacc
     3841 agacaaccat tacttatcct atcaatctgc cttatccaaa gatccaaacg aaaagagaga
     3901 ccacatggtc ttgttagaat ttgttactgc tgctggtatt acccatggta ttgatgaatt
     3961 gtacaaagca tccaacttga acatttcgag aaagcttacc atatcaaccc catcatgctc
     4021 tttcgaaaat tcaaatagca catccattcc ttcgcccgct tcctcatctc aaagccacac
     4081 tccaatgaga aacatgagct cactctctga taacagcgtt ttcagccgga atatggaaca
     4141 atcatcacca atcactccaa gtatgtacca atttggtcag cagcagtcaa acagtatatg
     4201 tggtagcacc gttagtgtga atagtctggt gaatacaaat aacaaacaaa ggatctacga
     4261 acaaatcacg ggtcctaaca gcaataacgc aaccaatgat tatattgatt tgctaaacct
     4321 aaatgagtct aacaaggaaa accaaaatcc cgcaacggcg cattacctca atgggggccc
     4381 acccaagaca agcttcatta accatggaat gttcccctcg ccaactggga ccataaatag
     4441 cggtaaatct agcagtgcct catctttaat ttcttttggt atgggcaata cccaagtaat
     4501 atagggcgcg ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa
     4561 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
     4621 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat
     4681 gaggtcgctc ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata
     4741 tagatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg acctcgaggg
     4801 ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     4861 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4921 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4981 agttgcgcag cctgaatggc gaatggcgcg acgcgccctg tagcggcgca ttaagcgcgg
     5041 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5101 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5161 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5221 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5281 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5341 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5401 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     5461 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatc
     5521 gacggtcgag gagaacttct agtatatcca catacctaat attattgcct tattaaaaat
     5581 ggaatcccaa caattacatc aaaatccaca ttctcttcaa aatcaattgt cctgtacttc
     5641 cttgttcatg tgtgttcaaa aacgttatat ttataggata attatactct atttctcaac
     5701 aagtaattgg ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat atcttgaccg
     5761 cagttaactg tgggaatact caggtatcgt aagatgcaag agttcgaatc tcttagcaac
     5821 cattattttt ttcctcaaca taacgagaac acacaggggc gctatcgcac agaatcaaat
     5881 tcgatgactg gaaatttttt gttaatttca gaggtcgcct gacgcatata cctttttcaa
     5941 ctgaaaaatt gggagaaaaa ggaaaggtga gaggccggaa ccggcttttc atatagaata
     6001 gagaagcgtt catgactaaa tgcttgcatc acaatacttg aagttgacaa tattatttaa
     6061 ggacctattg ttttttccaa taggtggtta gcaatcgtct tactttctaa cttttcttac
     6121 cttttacatt tcagcaatat atatatatat ttcaaggata taccattcta atgtctgccc
     6181 ctatgtctgc ccctaagaag atcgtcgttt tgccaggtga ccacgttggt caagaaatca
     6241 cagccgaagc cattaaggtt cttaaagcta tttctgatgt tcgttccaat gtcaagttcg
     6301 atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc tacaggtgtc ccacttccag
     6361 atgaggcgct ggaagcctcc aagaaggttg atgccgtttt gttaggtgct gtgggtggtc
     6421 ctaaatgggg taccggtagt gttagacctg aacaaggttt actaaaaatc cgtaaagaac
     6481 ttcaattgta cgccaactta agaccatgta actttgcatc cgactctctt ttagacttat
     6541 ctccaatcaa gccacaattt gctaaaggta ctgacttcgt tgttgtcaga gaattagtgg
     6601 gaggtattta ctttggtaag agaaaggaag acgatggtga tggtgtcgct tgggatagtg
     6661 aacaatacac cgttccagaa gtgcaaagaa tcacaagaat ggccgctttc atggccctac
     6721 aacatgagcc accattgcct atttggtcct tggataaagc taatgttttg gcctcttcaa
     6781 gattatggag aaaaactgtg gaggaaacca tcaagaacga attccctaca ttgaaggttc
     6841 aacatcaatt gattgattct gccgccatga tcctagttaa gaacccaacc cacctaaatg
     6901 gtattataat caccagcaac atgtttggtg atatcatctc cgatgaagcc tccgttatcc
     6961 caggttcctt gggtttgttg ccatctgcgt ccttggcctc tttgccagac aagaacaccg
     7021 catttggttt gtacgaacca tgccacggtt ctgctccaga tttgccaaag aataaggttg
     7081 accctatcgc cactatcttg tctgctgcaa tgatgttgaa attgtcattg aacttgcctg
     7141 aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt ggatgcaggt atcagaactg
     7201 gtgatttagg tggttccaac agtaccaccg aagtcggtga tgctgtcgcc gaagaagtta
     7261 agaaaatcct tgcttaaaaa gattctcttt ttttatgata tttgtacata aactttataa
     7321 atgaaattca taatagaaac gacacgaaat tacaaaatgg aatatgttca tagggtagac
     7381 gaaactatat acgcaatcta catacattta tcaagaagga gaaaaaggag gatagtaaag
     7441 gaatacaggt aagcaaattg atactaatgg ctcaacgtga taaggaaaaa gaattgcact
     7501 ttaacattaa tattgacaag gaggagggca ccacacaaaa agttaggtgt aacagaaaat
     7561 catgaaacta cgattcctaa tttgatattg gaggattttc tctaaaaaaa aaaaaataca
     7621 acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa taccttcttg
     7681 aaccatttcc cataatggtg aaagttccct caagaatttt actctgtcag aaacggcctt
     7741 acgacgtagt cgatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     7801 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     7861 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     7921 tcatcaccga aacgcgcga
//
