LOCUS       sequence-236954-        7686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of yeast codon-optimized mVenus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7686)
  AUTHORS   Botman D, de Groot DH, Schmidt P, Goedhart J, Teusink B
  TITLE     In vivo characterisation of fluorescent proteins in budding yeast.
  JOURNAL   Sci Rep. 2019 Feb 19;9(1):2234. doi: 10.1038/s41598-019-38913-z.
  PUBMED    30783202
REFERENCE   2  (bases 1 to 7686)
  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: "Sci Rep."; 
            date: "2019-02-19"; volume: "9(1):2234. doi"; pages: " 
            10.1038/s41598-019-38913-z"
COMMENT     Sequence Label: pDRF1-GW ymVenus
FEATURES             Location/Qualifiers
     source          1..7686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    800..824
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             851..1567
                     /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
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     protein_bind    complement(1576..1600)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1662..1681)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1663..1681)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1829..2016
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1884..1908)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2028..2045)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2028..2044)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2037..2059)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2480..3826
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3937..4740)
                     /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(4741..4956)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4981..4999)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5067..5171
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5172..5240,5241..6032)
                     /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(5390..5409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6203..6791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6692..6711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6945..6962
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7079..7100
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7115..7120,7121..7138,7139..7145)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7153..7169
                     /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     7158..7180
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7177..7193
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7177..7193
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7223..7678)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7360..7381)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7572..7591
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caagcttcct gaaacggaga aacataaaca ggcattgctg ggatcaccca tacatcactc
       61 tgttttgcct gaccttttcc ggtaatttga aaacaaaccc ggtctcgaag cggagatccg
      121 gcgataatta ccgcagaaat aaacccatac acgagacgta gaaccagccg cacatggccg
      181 gagaaactcc tgcgagaatt tcgtaaactc gcgcgcattg catctgtatt tcctaatgcg
      241 gcacttccag gcctcgatcg agaccgttta tccattgctt ttttgttgtc tttttccctc
      301 gttcacagaa agtctgaaga agctatagta gaactatgag ctttttttgt ttctgttttc
      361 cttttttttt tttttacctc tgtggaaatt gttactctca cactctttag ttcgtttgtt
      421 tgttttgttt attccaatta tgaccggtga cgaaacgtgg tcgatggtgg gtaccgctta
      481 tgctcccctc cattagtttc gattatataa aaaggccaaa tattgtatta ttttcaaatg
      541 tcctatcatt atcgtctaac atctaatttc tcttaaattt tttctctttc tttcctataa
      601 caccaatagt gaaaatcttt ttttcttcta tatctacaaa aacttttttt ttctatcaac
      661 ctcgttgata aattttttct ttaacaatcg ttaataatta attaattgga aaataaccat
      721 tttttctctc ttttatacac acattcaaaa gaaagaaaaa aaatataccc cagcctcgac
      781 tagtgaattc gcggccatca caagtttgta caaaaaagca ggctgctagc actagtaagc
      841 ttttaattaa atgtctaaag gtgaagaatt attcactggt gttgtcccaa ttttggttga
      901 attagatggt gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc
      961 tacttacggt aaattgacct taaaattgat ttgtactact ggtaaattgc cagttccatg
     1021 gccaacctta gtcactactt taggttatgg tttgcaatgt tttgctagat acccagatca
     1081 tatgaaacaa catgactttt tcaagtctgc catgccagaa ggttatgttc aagaaagaac
     1141 tatttttttc aaagatgacg gtaactacaa gaccagagct gaagtcaagt ttgaaggtga
     1201 taccttagtt aatagaatcg aattaaaagg tattgatttt aaagaagatg gtaacatttt
     1261 aggtcacaaa ttggaataca actataactc tcacaatgtt tacatcactg ctgacaaaca
     1321 aaagaatggt atcaaagcta acttcaaaat tagacacaac attgaagatg gtggtgttca
     1381 attagctgac cattatcaac aaaatactcc aattggtgat ggtccagtct tgttaccaga
     1441 caaccattac ttatcctatc aatctaaatt atccaaagat ccaaacgaaa agagagacca
     1501 catggtcttg ttagaatttg ttactgctgc tggtattacc catggtatgg atgaattgta
     1561 caaataagcg gccgcaccca gctttcttgt acaaagtggt gtagttaatt catgatggcc
     1621 gctgcaggtc gacctcgagg gggggcccgg tacccaattc gccctatagt gagtcgtatt
     1681 acgcgcggat ccagctttgg acttcttcgc cagaggtttg gtcaagtctc caatcaaggt
     1741 tgtcggcttg tctaccttgc cagaaattta cgaaaagatg gaaaagggtc aaatcgttgg
     1801 tagatacgtt gttgacactt ctaaataagc gaatttctta tgatttatga tttttattat
     1861 taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa
     1921 acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata
     1981 gcatgaggtc gctcttattg accacacctc taccggcatg ccaattcact ggccgtcgtt
     2041 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     2101 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     2161 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     2221 ggtatttcac accgcataat cggatcgtac ttgttaccca tcattgaatt ttgaacatcc
     2281 gaacctggga gttttccctg aaacagatag tatatttgaa cctgtataat aatatatagt
     2341 ctagcgcttt acggaagaca atgtatgtat ttcggttcct ggagaaacta ttgcatctat
     2401 tgcataggta atcttgcacg tcgcatcccc ggttcatttt ctgcgtttcc atcttgcact
     2461 tcaatagcat atctttgtta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa
     2521 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     2581 caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa
     2641 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     2701 aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac
     2761 aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc
     2821 ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt
     2881 tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc
     2941 ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc
     3001 gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg
     3061 atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta
     3121 cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac
     3181 tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag
     3241 tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca
     3301 gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt
     3361 ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc ttcagagcgc
     3421 ttttggtttt caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc
     3481 ggaataggaa cttcaaagcg tttccgaaaa cgagcgcttc cgaaaatgca acgcgagctg
     3541 cgcacataca gctcactgtt cacgtcgcac ctatatctgc gtgttgcctg tatatatata
     3601 tacatgagaa gaacggcata gtgcgtgttt atgcttaaat gcgtacttat atgcgtctat
     3661 ttatgtagga tgaaaggtag tctagtacct cctgtgatat tatcccattc catgcggggt
     3721 atcgtatgct tccttcagca ctacccttta gctgttctat atgctgccac tcctcaattg
     3781 gattagtctc atccttcaat gctatcattt cctttgatat tggatcgatc cgatgataag
     3841 ctgtcaaaca tgagaattgg gtaataactg atataattaa attgaagctc taatttgtga
     3901 gtttagtata catgcattta cttataatac agttttttag ttttgctggc cgcatcttct
     3961 caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc
     4021 ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc
     4081 cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt
     4141 cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc
     4201 gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt
     4261 agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt
     4321 tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc
     4381 attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac
     4441 tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga
     4501 taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg
     4561 tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt
     4621 ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag
     4681 cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat
     4741 gatttatctt cgtttcctgc atgtttttgt tctgtgcagt tgggttaaga atactgggca
     4801 atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt
     4861 ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa
     4921 tcaaaaaaaa gaataaaaaa aaaatgatga attgaaaagc taattcttga agacgaaagg
     4981 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     5041 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5101 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5161 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5221 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5281 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5341 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5401 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5461 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5521 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5581 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5641 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5701 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5761 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5821 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5881 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5941 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6001 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6061 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6121 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6181 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6241 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6301 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6361 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6421 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6481 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6541 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6601 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6661 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6721 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6781 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6841 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6901 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6961 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     7021 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     7081 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     7141 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     7201 acgccaagct taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     7261 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     7321 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     7381 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     7441 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     7501 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     7561 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     7621 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     7681 attcgc
//
