LOCUS       sequence-407652-        8440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ymTq2-E2A-ymScarletl in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8440)
  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 8440)
  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-E2A-ymScarletl
FEATURES             Location/Qualifiers
     source          1..8440
                     /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(2330..2354)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2416..2435)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2417..2435)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2583..2770
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2638..2662)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2782..2799)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2782..2798)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2791..2813)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3234..4580
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4691..5494)
                     /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(5495..5710)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5735..5753)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5821..5925
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5926..5994,5995..6786)
                     /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(6144..6163)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6957..7545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7446..7465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7699..7716
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7833..7854
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7869..7874,7875..7892,7893..7899)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7907..7923
                     /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     7912..7934
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7931..7947
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7931..7947
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7977..8432)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8114..8135)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8326..8345
                     /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 gtataaaggt tctggtggcg
     1561 ctaccaattt ctcactacta aaactggcgg gtgacgtcga attgaatccc ggccctcccg
     1621 ggatggttag taaaggtgaa gctgttataa aagaattcat gaggtttaaa gttcatatgg
     1681 aaggttcaat gaatggtcat gaatttgaaa ttgaaggtga aggtgaaggt agaccatatg
     1741 aaggtacaca aactgctaaa ttgaaagtta ctaaaggtgg tccattacca ttttcttggg
     1801 atattttgtc tccacaattc atgtatggtt ctagagcttt tattaagcat ccagctgata
     1861 ttccagatta ttataaacaa tcttttcctg aaggttttaa atgggaaaga gttatgaatt
     1921 tcgaagatgg tggtgctgtt actgttactc aagatacttc tttggaagat ggtactttaa
     1981 tctataaagt taaattgaga ggtactaatt ttccaccaga tggtccagtt atgcaaaaga
     2041 aaactatggg ttgggaagca tctactgaaa gattgtatcc agaagatggt gttttgaaag
     2101 gagacattaa gatggctttg agattgaaag atggtggtcg gtacctggcc gacttcaaga
     2161 ccacctataa agctaaaaaa ccagttcaaa tgccaggtgc atataatgtt gatagaaagt
     2221 tagatataac gtcgcataac gaggactata ctgtagttga acaatatgaa cgtagtgaag
     2281 gtagacatag taccggagga atggatgaat tgtataaata agcggccgca cccagctttc
     2341 ttgtacaaag tggtgtagtt aattcatgat ggccgctgca ggtcgacctc gagggggggc
     2401 ccggtaccca attcgcccta tagtgagtcg tattacgcgc ggatccagct ttggacttct
     2461 tcgccagagg tttggtcaag tctccaatca aggttgtcgg cttgtctacc ttgccagaaa
     2521 tttacgaaaa gatggaaaag ggtcaaatcg ttggtagata cgttgttgac acttctaaat
     2581 aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg
     2641 tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact
     2701 ctttcctgta ggtcaggttg ctttctcagg tatagcatga ggtcgctctt attgaccaca
     2761 cctctaccgg catgccaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     2821 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     2881 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     2941 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca taatcggatc
     3001 gtacttgtta cccatcattg aattttgaac atccgaacct gggagttttc cctgaaacag
     3061 atagtatatt tgaacctgta taataatata tagtctagcg ctttacggaa gacaatgtat
     3121 gtatttcggt tcctggagaa actattgcat ctattgcata ggtaatcttg cacgtcgcat
     3181 ccccggttca ttttctgcgt ttccatcttg cacttcaata gcatatcttt gttaacgaag
     3241 catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     3301 agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac
     3361 gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca
     3421 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta
     3481 ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt
     3541 ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc
     3601 ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat
     3661 tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc
     3721 tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt
     3781 gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta
     3841 tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt
     3901 attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat
     3961 actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa
     4021 ggtggatggg taggttatat agggatatag cacagagata tatagcaaag agatactttt
     4081 gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg
     4141 tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag cgctctgaag
     4201 ttcctatact ttctagctag agaataggaa cttcggaata ggaacttcaa agcgtttccg
     4261 aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca tacagctcac tgttcacgtc
     4321 gcacctatat ctgcgtgttg cctgtatata tatatacatg agaagaacgg catagtgcgt
     4381 gtttatgctt aaatgcgtac ttatatgcgt ctatttatgt aggatgaaag gtagtctagt
     4441 acctcctgtg atattatccc attccatgcg gggtatcgta tgcttccttc agcactaccc
     4501 tttagctgtt ctatatgctg ccactcctca attggattag tctcatcctt caatgctatc
     4561 atttcctttg atattggatc gatccgatga taagctgtca aacatgagaa ttgggtaata
     4621 actgatataa ttaaattgaa gctctaattt gtgagtttag tatacatgca tttacttata
     4681 atacagtttt ttagttttgc tggccgcatc ttctcaaata tgcttcccag cctgcttttc
     4741 tgtaacgttc accctctacc ttagcatccc ttccctttgc aaatagtcct cttccaacaa
     4801 taataatgtc agatcctgta gagaccacat catccacggt tctatactgt tgacccaatg
     4861 cgtctccctt gtcatctaaa cccacaccgg gtgtcataat caaccaatcg taaccttcat
     4921 ctcttccacc catgtctctt tgagcaataa agccgataac aaaatctttg tcgctcttcg
     4981 caatgtcaac agtaccctta gtatattctc cagtagatag ggagcccttg catgacaatt
     5041 ctgctaacat caaaaggcct ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac
     5101 cgctaacaat acctgggccc accacaccgt gtgcattcgt aatgtctgcc cattctgcta
     5161 ttctgtatac acccgcagag tactgcaatt tgactgtatt accaatgtca gcaaattttc
     5221 tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc ttaactgtgc
     5281 cctccatgga aaaatcagtc aagatatcca catgtgtttt tagtaaacaa attttgggac
     5341 ctaatgcttc aactaactcc agtaattcct tggtggtacg aacatccaat gaagcacaca
     5401 agtttgtttg cttttcgtgc atgatattaa atagcttggc agcaacagga ctaggatgag
     5461 tagcagcacg ttccttatat gtagctttcg acatgattta tcttcgtttc ctgcatgttt
     5521 ttgttctgtg cagttgggtt aagaatactg ggcaatttca tgtttcttca acactacata
     5581 tgcgtatata taccaatcta agtctgtgct ccttccttcg ttcttccttc tgttcggaga
     5641 ttaccgaatc aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa aaaaaaaatg
     5701 atgaattgaa aagctaattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     5761 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5821 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5881 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5941 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6001 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6061 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6121 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6181 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6241 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6301 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6361 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6421 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6481 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6541 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6601 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6661 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6721 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6781 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6841 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6901 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6961 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7021 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7081 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7141 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7201 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7261 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7321 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7381 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7441 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7501 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7561 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7621 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7681 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     7741 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     7801 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     7861 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     7921 caatttcaca caggaaacag ctatgaccat gattacgcca agcttaccgc atcaggaaat
     7981 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     8041 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     8101 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     8161 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     8221 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     8281 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     8341 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     8401 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc
//
