LOCUS       sequence-407704-        8407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ymTq2-ymScarletl in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8407)
  AUTHORS   Botman D, de Groot DH, Wehrens M, van Rossum K, Yska S, van der 
            Kieft B, Albert V, Minghetti N, Goedhart J, Teusink B
  TITLE     Side-by-side systematic characterization of novel FPs in budding 
            yeast
  JOURNAL   bioRxiv 2025.06.27.661975; doi: 
            https://doi.org/10.1101/2025.06.27.661975
REFERENCE   2  (bases 1 to 8407)
  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: "bioRxiv 
            2025.06.27.661975; doi: https://doi.org/10.1101/2025.06.27.661975"
COMMENT     Sequence Label: pDRF1-GW ymTq2-ymScarletI
FEATURES             Location/Qualifiers
     source          1..8407
                     /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(2297..2321)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2383..2402)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2384..2402)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2550..2737
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2605..2629)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2749..2766)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2749..2765)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2758..2780)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3201..4547
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4658..5461)
                     /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(5462..5677)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5702..5720)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5788..5892
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5893..5961,5962..6753)
                     /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(6111..6130)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6924..7512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7413..7432
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7666..7683
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7800..7821
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7836..7841,7842..7859,7860..7866)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7874..7890
                     /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     7879..7901
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7898..7914
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7898..7914
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7944..8399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8081..8102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8293..8312
                     /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 atggttagta
      841 aaggtgaaga attgttcact ggtgtagtgc caattttagt agaattagat ggtgacgtta
      901 acggtcataa gttctctgta agcggtgaag gtgaaggtga cgcaacatac ggtaaattaa
      961 ctttgaaatt catttgtact acaggtaaat taccagttcc atggccaact ttggttacta
     1021 ctttgtcttg gggtgttcaa tgtttcgcta gatacccaga tcatatgaag caacatgatt
     1081 tctttaagtc tgctatgcca gaaggttacg ttcaagaaag aactatcttc ttcaaggatg
     1141 atggtaacta taagactaga gctgaagtta agttcgaagg agatacttta gttaatagaa
     1201 ttgaattgaa gggtattgat tttaaggaag atggtaatat tttgggtcat aagttggaat
     1261 ataattactt ctctgataac gtttacatta ctgctgataa gcaaaagaac ggtattaagg
     1321 cgaacttcaa gattagacat aacattgaag atggtggtgt tcaattagct gatcattacc
     1381 aacaaaacac tccaattggt gacggtccag ttttgttgcc agataaccat tacttgtcta
     1441 ctcaatctaa gttgtctaag gatccaaacg aaaagagaga tcatatggtt ttgttggaat
     1501 tcgttactgc tgctgggatc actttaggta tggatgaatt gtataaatcc ggactcagat
     1561 cttcggatcc accggtcgcc acccccggga tggttagtaa aggtgaagct gttataaaag
     1621 aattcatgag gtttaaagtt catatggaag gttcaatgaa tggtcatgaa tttgaaattg
     1681 aaggtgaagg tgaaggtaga ccatatgaag gtacacaaac tgctaaattg aaagttacta
     1741 aaggtggtcc attaccattt tcttgggata ttttgtctcc acaattcatg tatggttcta
     1801 gagcttttat taagcatcca gctgatattc cagattatta taaacaatct tttcctgaag
     1861 gttttaaatg ggaaagagtt atgaatttcg aagatggtgg tgctgttact gttactcaag
     1921 atacttcttt ggaagatggt actttaatct ataaagttaa attgagaggt actaattttc
     1981 caccagatgg tccagttatg caaaagaaaa ctatgggttg ggaagcatct actgaaagat
     2041 tgtatccaga agatggtgtt ttgaaaggag acattaagat ggctttgaga ttgaaagatg
     2101 gtggtcggta cctggccgac ttcaagacca cctataaagc taaaaaacca gttcaaatgc
     2161 caggtgcata taatgttgat agaaagttag atataacgtc gcataacgag gactatactg
     2221 tagttgaaca atatgaacgt agtgaaggta gacatagtac cggaggaatg gatgaattgt
     2281 ataaataagc ggccgcaccc agctttcttg tacaaagtgg tgtagttaat tcatgatggc
     2341 cgctgcaggt cgacctcgag ggggggcccg gtacccaatt cgccctatag tgagtcgtat
     2401 tacgcgcgga tccagctttg gacttcttcg ccagaggttt ggtcaagtct ccaatcaagg
     2461 ttgtcggctt gtctaccttg ccagaaattt acgaaaagat ggaaaagggt caaatcgttg
     2521 gtagatacgt tgttgacact tctaaataag cgaatttctt atgatttatg atttttatta
     2581 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
     2641 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     2701 agcatgaggt cgctcttatt gaccacacct ctaccggcat gccaattcac tggccgtcgt
     2761 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     2821 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     2881 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     2941 cggtatttca caccgcataa tcggatcgta cttgttaccc atcattgaat tttgaacatc
     3001 cgaacctggg agttttccct gaaacagata gtatatttga acctgtataa taatatatag
     3061 tctagcgctt tacggaagac aatgtatgta tttcggttcc tggagaaact attgcatcta
     3121 ttgcataggt aatcttgcac gtcgcatccc cggttcattt tctgcgtttc catcttgcac
     3181 ttcaatagca tatctttgtt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca
     3241 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     3301 gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa
     3361 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     3421 gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta
     3481 caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct
     3541 cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg
     3601 ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt
     3661 cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc
     3721 cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt
     3781 gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact
     3841 acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta
     3901 ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga
     3961 gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac
     4021 agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat
     4081 tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg
     4141 cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt
     4201 cggaatagga acttcaaagc gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct
     4261 gcgcacatac agctcactgt tcacgtcgca cctatatctg cgtgttgcct gtatatatat
     4321 atacatgaga agaacggcat agtgcgtgtt tatgcttaaa tgcgtactta tatgcgtcta
     4381 tttatgtagg atgaaaggta gtctagtacc tcctgtgata ttatcccatt ccatgcgggg
     4441 tatcgtatgc ttccttcagc actacccttt agctgttcta tatgctgcca ctcctcaatt
     4501 ggattagtct catccttcaa tgctatcatt tcctttgata ttggatcgat ccgatgataa
     4561 gctgtcaaac atgagaattg ggtaataact gatataatta aattgaagct ctaatttgtg
     4621 agtttagtat acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc
     4681 tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc
     4741 cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat
     4801 ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg
     4861 tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc
     4921 cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag
     4981 tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg
     5041 ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg
     5101 cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga
     5161 ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg
     5221 ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat
     5281 gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg
     5341 tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata
     5401 gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca
     5461 tgatttatct tcgtttcctg catgtttttg ttctgtgcag ttgggttaag aatactgggc
     5521 aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct
     5581 tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa
     5641 atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag ctaattcttg aagacgaaag
     5701 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5761 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5821 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5881 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5941 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6001 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6061 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6121 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6181 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6241 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6301 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6361 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6421 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6481 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6541 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6601 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6661 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6721 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6781 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6841 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6901 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6961 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7021 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7081 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7141 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7201 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7261 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7321 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7381 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7441 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7501 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7561 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7621 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7681 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7741 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     7801 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     7861 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     7921 tacgccaagc ttaccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     7981 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     8041 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     8101 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     8161 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     8221 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     8281 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     8341 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     8401 cattcgc
//
