LOCUS       sequence-437598-        8099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of Cyterm-ymiRFP680 in budding yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8099)
  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 8099)
  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 Cyterm-ymiRFP680
FEATURES             Location/Qualifiers
     source          1..8099
                     /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"
     regulatory      1024..1033
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1989..2013)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2075..2094)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2076..2094)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2242..2429
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2297..2321)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2441..2458)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2441..2457)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2450..2472)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2893..4239
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4350..5153)
                     /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(5154..5369)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5394..5412)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5480..5584
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5585..5653,5654..6445)
                     /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(5803..5822)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6616..7204
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7105..7124
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7358..7375
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7492..7513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7528..7533,7534..7551,7552..7558)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7566..7582
                     /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     7571..7593
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7590..7606
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7590..7606
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7636..8091)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7773..7794)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7985..8004
                     /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 gctaccggac
      841 tcagatctgg taccccttgc agcttccagt tgtcaggaag agaaggctcc catggatcct
      901 gtggtggtgc tggggctctg tctctcctgt ttgcttctcc tttcactctg gaaacagagc
      961 tatgggggag ggaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     1021 gtcgccacca tggctgaagg ttctgttgct agacaaccag atttgttgac ttgtgatgat
     1081 gaaccaattc atattccagg tgctattcaa ccacatggtt tgttattagc tttggctgct
     1141 gatatgacca ttgttgctgg ttctgataac ttgcctgaat tgactggttt agctattggt
     1201 gccttgatcg gtagatctgc tgctgatgtt ttcgattctg aaacccataa tagattgact
     1261 attgctttgg ctgaaccagg tgctgctgtc ggtgctccaa ttaccgtcgg tttcaccatg
     1321 cgtaaggatg ccggtttcat tggttcttgg catagacatg atcaattaat tttcttggaa
     1381 ttggaaccac cacaacgtga cgttgccgaa ccacaggctt tcttccgtag aaccaactct
     1441 gctattagaa gattacaagc tgctgaaact ttggaatctg cttgtgctgc tgctgctcaa
     1501 gaagttcgta agattactgg tttcgataga gttatgatct acagatttgc ttctgatttc
     1561 tctggtgaag ttattgctga agatagatgt gctgaagttg aatctaagtt gggtttgcat
     1621 tacccagctt ctactgttcc agctcaagct agaagattgt acactattaa tccagttaga
     1681 attattccag atatcaacta cagaccagtc ccagttaccc cagatttgaa cccagttact
     1741 ggtagaccta ttgatttatc tttcgctatt ttgagatctg tttccccatg tcatttggaa
     1801 ttcatgagaa acatcggtat gcatggtacc atgtctattt ctattttgag aggtgaaaga
     1861 ttgtggggtt tgattgtttg tcatcataga actccatact acgttgactt ggatggtaga
     1921 caagcctgta agagagttgc tgaaagattg gctacccaaa ttggtgttat ggaagaataa
     1981 gcggccgcac ccagctttct tgtacaaagt ggtgtagtta attcatgatg gccgctgcag
     2041 gtcgacctcg agggggggcc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg
     2101 gatccagctt tggacttctt cgccagaggt ttggtcaagt ctccaatcaa ggttgtcggc
     2161 ttgtctacct tgccagaaat ttacgaaaag atggaaaagg gtcaaatcgt tggtagatac
     2221 gttgttgaca cttctaaata agcgaatttc ttatgattta tgatttttat tattaaataa
     2281 gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaaa
     2341 ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagcatgag
     2401 gtcgctctta ttgaccacac ctctaccggc atgccaattc actggccgtc gttttacaac
     2461 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     2521 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     2581 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2641 cacaccgcat aatcggatcg tacttgttac ccatcattga attttgaaca tccgaacctg
     2701 ggagttttcc ctgaaacaga tagtatattt gaacctgtat aataatatat agtctagcgc
     2761 tttacggaag acaatgtatg tatttcggtt cctggagaaa ctattgcatc tattgcatag
     2821 gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt tccatcttgc acttcaatag
     2881 catatctttg ttaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga
     2941 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     3001 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg
     3061 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     3121 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg
     3181 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg
     3241 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga
     3301 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt
     3361 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata
     3421 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc
     3481 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag
     3541 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt
     3601 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat
     3661 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat
     3721 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag
     3781 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag cgcttttggt
     3841 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcggaatag
     3901 gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat
     3961 acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga
     4021 gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta
     4081 ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat
     4141 gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt
     4201 ctcatccttc aatgctatca tttcctttga tattggatcg atccgatgat aagctgtcaa
     4261 acatgagaat tgggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt
     4321 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat
     4381 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca
     4441 aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt
     4501 ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc
     4561 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca
     4621 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg
     4681 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct
     4741 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta
     4801 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta
     4861 ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc
     4921 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt
     4981 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga
     5041 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca
     5101 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat
     5161 cttcgtttcc tgcatgtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat
     5221 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt
     5281 tcttccttct gttcggagat taccgaatca aaaaaatttc aaggaaaccg aaatcaaaaa
     5341 aaagaataaa aaaaaaatga tgaattgaaa agctaattct tgaagacgaa agggcctcgt
     5401 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     5461 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5521 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5581 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5641 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5701 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5761 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5821 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5881 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5941 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6001 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6061 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6121 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6181 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6241 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6301 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6361 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6421 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6481 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6541 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6601 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6661 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6721 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6781 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6841 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6901 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6961 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7021 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7081 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7141 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7201 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7261 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     7321 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     7381 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     7441 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     7501 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     7561 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     7621 gcttaccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     7681 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     7741 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     7801 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     7861 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     7921 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     7981 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     8041 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgc
//
