LOCUS       sequence-223815-        7668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of yeast codon-optimized mScarletI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7668)
  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 7668)
  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 ymScarletI
FEATURES             Location/Qualifiers
     source          1..7668
                     /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(1558..1582)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1644..1663)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1645..1663)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1811..1998
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1866..1890)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2010..2027)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2010..2026)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2019..2041)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2462..3808
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3919..4722)
                     /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(4723..4938)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4963..4981)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5049..5153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5154..5222,5223..6014)
                     /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(5372..5391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6185..6773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6674..6693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6927..6944
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7061..7082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7097..7102,7103..7120,7121..7127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7135..7151
                     /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     7140..7162
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7159..7175
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7159..7175
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7205..7660)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7342..7363)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7554..7573
                     /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 atggttagta aaggtgaagc tgttataaaa gaattcatga ggtttaaagt
      901 tcatatggaa ggttcaatga atggtcatga atttgaaatt gaaggtgaag gtgaaggtag
      961 accatatgaa ggtacacaaa ctgctaaatt gaaagttact aaaggtggtc cattaccatt
     1021 ttcttgggat attttgtctc cacaattcat gtatggttct agagctttta ttaagcatcc
     1081 agctgatatt ccagattatt ataaacaatc ttttcctgaa ggttttaaat gggaaagagt
     1141 tatgaatttc gaagatggtg gtgctgttac tgttactcaa gatacttctt tggaagatgg
     1201 tactttaatc tataaagtta aattgagagg tactaatttt ccaccagatg gtccagttat
     1261 gcaaaagaaa actatgggtt gggaagcatc tactgaaaga ttgtatccag aagatggtgt
     1321 tttgaaagga gacattaaga tggctttgag attgaaagat ggtggtcggt acctggccga
     1381 cttcaagacc acctataaag ctaaaaaacc agttcaaatg ccaggtgcat ataatgttga
     1441 tagaaagtta gatataacgt cgcataacga ggactatact gtagttgaac aatatgaacg
     1501 tagtgaaggt agacatagta ccggaggaat ggatgaattg tataaataag cggccgcacc
     1561 cagctttctt gtacaaagtg gtgtagttaa ttcatgatgg ccgctgcagg tcgacctcga
     1621 gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgg atccagcttt
     1681 ggacttcttc gccagaggtt tggtcaagtc tccaatcaag gttgtcggct tgtctacctt
     1741 gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt ggtagatacg ttgttgacac
     1801 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
     1861 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacganaat tcttattctt
     1921 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg tcgctcttat
     1981 tgaccacacc tctaccggca tgccaattca ctggccgtcg ttttacaacg tcgtgactgg
     2041 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     2101 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2161 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2221 atcggatcgt acttgttacc catcattgaa ttttgaacat ccgaacctgg gagttttccc
     2281 tgaaacagat agtatatttg aacctgtata ataatatata gtctagcgct ttacggaaga
     2341 caatgtatgt atttcggttc ctggagaaac tattgcatct attgcatagg taatcttgca
     2401 cgtcgcatcc ccggttcatt ttctgcgttt ccatcttgca cttcaatagc atatctttgt
     2461 taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
     2521 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
     2581 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
     2641 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
     2701 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
     2761 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
     2821 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
     2881 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
     2941 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
     3001 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
     3061 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
     3121 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
     3181 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
     3241 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
     3301 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
     3361 ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg
     3421 ctctgaagtt cctatacttt ctagctagag aataggaact tcggaatagg aacttcaaag
     3481 cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc tgcgcacata cagctcactg
     3541 ttcacgtcgc acctatatct gcgtgttgcc tgtatatata tatacatgag aagaacggca
     3601 tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct atttatgtag gatgaaaggt
     3661 agtctagtac ctcctgtgat attatcccat tccatgcggg gtatcgtatg cttccttcag
     3721 cactaccctt tagctgttct atatgctgcc actcctcaat tggattagtc tcatccttca
     3781 atgctatcat ttcctttgat attggatcga tccgatgata agctgtcaaa catgagaatt
     3841 gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta tacatgcatt
     3901 tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg cttcccagcc
     3961 tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa atagtcctct
     4021 tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc tatactgttg
     4081 acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca accaatcgta
     4141 accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa aatctttgtc
     4201 gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg agcccttgca
     4261 tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt ctgccgcctg
     4321 cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca
     4381 ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac caatgtcagc
     4441 aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct ttagcggctt
     4501 aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta gtaaacaaat
     4561 tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa catccaatga
     4621 agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag caacaggact
     4681 aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc ttcgtttcct
     4741 gcatgttttt gttctgtgca gttgggttaa gaatactggg caatttcatg tttcttcaac
     4801 actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt cttccttctg
     4861 ttcggagatt accgaatcaa aaaaatttca aggaaaccga aatcaaaaaa aagaataaaa
     4921 aaaaaatgat gaattgaaaa gctaattctt gaagacgaaa gggcctcgtg atacgcctat
     4981 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5041 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5101 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5161 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5221 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5281 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5341 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5401 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5461 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5521 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5581 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5641 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5701 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5761 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5821 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5881 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5941 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6001 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6061 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6121 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6181 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6241 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6301 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6361 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6421 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6481 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6541 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6601 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6661 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6721 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6781 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6841 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6901 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     6961 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     7021 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     7081 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     7141 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cttaccgcat
     7201 caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     7261 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     7321 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     7381 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     7441 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     7501 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     7561 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     7621 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgc
//
