LOCUS       sequence-223818-        7689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of ymYPET.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7689)
  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 7689)
  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 ymYPET
FEATURES             Location/Qualifiers
     source          1..7689
                     /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"
     protein_bind    complement(1579..1603)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1665..1684)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1666..1684)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1832..2019
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1887..1911)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2031..2048)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2031..2047)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2040..2062)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2483..3829
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3940..4743)
                     /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(4744..4959)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4984..5002)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5070..5174
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5175..5243,5244..6035)
                     /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(5393..5412)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6206..6794
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6695..6714
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6948..6965
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7082..7103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7118..7123,7124..7141,7142..7148)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7156..7172
                     /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     7161..7183
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7180..7196
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7180..7196
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7226..7681)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7363..7384)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7575..7594
                     /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 atggtctcca agggcgaaga attgttcacc ggtgtggtcc caatcttggt
      901 cgagttggac ggtgacgtca acggtcacaa gttctctgtt tccggtgaag gtgaaggaga
      961 cgccacctac ggtaaattga ccttgaagtt gttgtgcacc accggcaagc tgccagttcc
     1021 atggccaaca ttggtcacca ctttaggtta tggtttacaa tgttttgcta gatatccaga
     1081 tcatatgaaa caacatgatt tcttcaaatc tgctatgcca gaaggttatg ttcaagaaag
     1141 aactatattt ttcaaagatg atggtaatta taaaactaga gctgaagtta aatttgaagg
     1201 agatacttta gttaatagaa ttgaattgaa aggtattgat tttaaagaag atggtaatat
     1261 tttaggtcat aaattagaat ataattataa ttctcataat gtttatatta cagctgataa
     1321 acaaaagaat ggtattaagg ctaattttaa aattagacat aatattgaag atggtggtgt
     1381 tcaattagct gatcattatc aacaaaatac tcctattggt gacggtccag ttttattacc
     1441 tgataatcat tatttgtcat atcaatctaa attgtcaaaa gatccaaatg aaaaaagaga
     1501 tcatatggtt ttgttagaat ttttgactgc agctggtatt actttaggta tggatgaatt
     1561 gtataaataa gcggccgcac ccagctttct tgtacaaagt ggtgtagtta attcatgatg
     1621 gccgctgcag gtcgacctcg agggggggcc cggtacccaa ttcgccctat agtgagtcgt
     1681 attacgcgcg gatccagctt tggacttctt cgccagaggt ttggtcaagt ctccaatcaa
     1741 ggttgtcggc ttgtctacct tgccagaaat ttacgaaaag atggaaaagg gtcaaatcgt
     1801 tggtagatac gttgttgaca cttctaaata agcgaatttc ttatgattta tgatttttat
     1861 tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt
     1921 aaaacganaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt
     1981 atagcatgag gtcgctctta ttgaccacac ctctaccggc atgccaattc actggccgtc
     2041 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2101 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2161 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     2221 tgcggtattt cacaccgcat aatcggatcg tacttgttac ccatcattga attttgaaca
     2281 tccgaacctg ggagttttcc ctgaaacaga tagtatattt gaacctgtat aataatatat
     2341 agtctagcgc tttacggaag acaatgtatg tatttcggtt cctggagaaa ctattgcatc
     2401 tattgcatag gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt tccatcttgc
     2461 acttcaatag catatctttg ttaacgaagc atctgtgctt cattttgtag aacaaaaatg
     2521 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     2581 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     2641 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2701 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     2761 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     2821 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     2881 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     2941 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     3001 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     3061 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     3121 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     3181 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     3241 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     3301 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     3361 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag
     3421 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     3481 ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag
     3541 ctgcgcacat acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat
     3601 atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc
     3661 tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg
     3721 ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa
     3781 ttggattagt ctcatccttc aatgctatca tttcctttga tattggatcg atccgatgat
     3841 aagctgtcaa acatgagaat tgggtaataa ctgatataat taaattgaag ctctaatttg
     3901 tgagtttagt atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct
     3961 tctcaaatat gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct
     4021 tccctttgca aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc
     4081 atccacggtt ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg
     4141 tgtcataatc aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa
     4201 gccgataaca aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc
     4261 agtagatagg gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt
     4321 tgttacttct tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg
     4381 tgcattcgta atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt
     4441 gactgtatta ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc
     4501 ggataatgcc tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac
     4561 atgtgttttt agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt
     4621 ggtggtacga acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa
     4681 tagcttggca gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga
     4741 catgatttat cttcgtttcc tgcatgtttt tgttctgtgc agttgggtta agaatactgg
     4801 gcaatttcat gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc
     4861 cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc aaggaaaccg
     4921 aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaaa agctaattct tgaagacgaa
     4981 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     5041 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5101 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5161 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5221 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5281 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5341 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5401 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5461 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5521 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5581 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5641 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5701 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5761 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5821 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5881 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5941 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6001 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6061 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6121 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6181 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6241 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6301 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6361 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6421 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6481 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6541 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6601 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6661 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6721 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6781 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6841 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6901 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6961 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     7021 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     7081 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     7141 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     7201 attacgccaa gcttaccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg
     7261 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     7321 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     7381 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     7441 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     7501 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     7561 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     7621 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     7681 tccattcgc
//
