LOCUS       sequence-437582-        8597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of Cyterm-ytd8oxStayGold in budding yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8597)
  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 8597)
  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-ytd8oxStayGold
FEATURES             Location/Qualifiers
     source          1..8597
                     /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(2487..2511)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2573..2592)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2574..2592)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2740..2927
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2795..2819)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2939..2956)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2939..2955)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2948..2970)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3391..4737
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4848..5651)
                     /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(5652..5867)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5892..5910)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5978..6082
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6083..6151,6152..6943)
                     /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(6301..6320)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7114..7702
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7603..7622
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7856..7873
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7990..8011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8026..8031,8032..8049,8050..8056)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8064..8080
                     /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     8069..8091
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8088..8104
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8088..8104
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(8134..8589)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8271..8292)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8483..8502
                     /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 tggtttctac tggtgaagaa ttgtttactg gtgttgttcc atttaagttc
     1081 caattgaagg gtaccattaa cggtaagtct ttcactgttg aaggtgaagg tgagggtaac
     1141 tctcatgaag gttctcataa gggtaagtat gtttgtactt ccggtaagtt gccaatgtct
     1201 tgggctgctc taggtacttc tttcggttat ggtatgaaat attacactaa atacccatct
     1261 ggtttaaaga attggttcca tgaagtcatg cctgaaggtt tcacttacga tagacatatt
     1321 caatataaag gtgatggttc tattcatgct aagcatcaac atttcatgaa gaatggtact
     1381 tatcacaaca ttgttgaatt cacaggtcaa gattttaaag aaaactctcc agttttgacc
     1441 ggtgatatga acgtttcttt gccaaacgaa gtccaacaca ttccaagaga cgatggtgtt
     1501 gaatgtccag ttactttgtt gtatccattg ttgtctgata aatctaagtg tgttgaggct
     1561 tatcaaaaca caattatcaa gccattgcat aatcaaccag cccctgatgt tccatatcac
     1621 tggatcagaa agcaatacac tcaatctaag gatgatactg aagaaagaga tcatattatc
     1681 caatctgaaa ccttagaagc tcatttgtac agtagaacta aattagaagg tcatggtact
     1741 ggttctactg gttctggttc ttctggtact gcttcctctg aagataataa tattgatatg
     1801 gttagcactg gtgaagaatt gttcactggt gttgttccat ttaagttcca attgaaaggt
     1861 accattaacg gtaagtcatt tacagtcgaa ggtgaaggtg aagggaactc tcatgaaggt
     1921 tcacacaaag gtaagtacgt ctgtacatcc ggtaagttgc ctatgagttg ggctgcttta
     1981 ggtacttctt tcggttacgg tatgaaatat tacactaaat acccatccgg tttgaagaat
     2041 tggttccatg aagttatgcc tgaaggtttc acttatgata gacacattca atacaagggt
     2101 gatggttcaa ttcacgctaa gcaccaacat tttatgaaaa acggtactta ccataacatt
     2161 gttgaattca ccggtcaaga ttttaaggaa aattctccag ttttaactgg tgatatgaac
     2221 gtctccttgc caaatgaagt tcaacacatc ccaagagatg atggtgttga atgtccagtc
     2281 accttgttgt atccattatt atctgataaa tctaagtgtg ttgaagccta ccaaaacact
     2341 ataattaagc cattgcacaa ccaaccagct cctgatgttc cataccattg gattagaaag
     2401 caatacactc aaagtaagga tgatactgaa gaaagagatc atattattca atctgaaact
     2461 ttggaagctc atttgtaagc ggccgcaccc agctttcttg tacaaagtgg tgtagttaat
     2521 tcatgatggc cgctgcaggt cgacctcgag ggggggcccg gtacccaatt cgccctatag
     2581 tgagtcgtat tacgcgcgga tccagctttg gacttcttcg ccagaggttt ggtcaagtct
     2641 ccaatcaagg ttgtcggctt gtctaccttg ccagaaattt acgaaaagat ggaaaagggt
     2701 caaatcgttg gtagatacgt tgttgacact tctaaataag cgaatttctt atgatttatg
     2761 atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct
     2821 taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt
     2881 tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat gccaattcac
     2941 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     3001 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     3061 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     3121 cgcatctgtg cggtatttca caccgcataa tcggatcgta cttgttaccc atcattgaat
     3181 tttgaacatc cgaacctggg agttttccct gaaacagata gtatatttga acctgtataa
     3241 taatatatag tctagcgctt tacggaagac aatgtatgta tttcggttcc tggagaaact
     3301 attgcatcta ttgcataggt aatcttgcac gtcgcatccc cggttcattt tctgcgtttc
     3361 catcttgcac ttcaatagca tatctttgtt aacgaagcat ctgtgcttca ttttgtagaa
     3421 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     3481 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg
     3541 taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     3601 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt
     3661 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta
     3721 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg
     3781 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct
     3841 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata
     3901 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag
     3961 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc
     4021 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga
     4081 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg
     4141 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg
     4201 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta
     4261 ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt
     4321 cttcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga
     4381 ataggaactt cggaatagga acttcaaagc gtttccgaaa acgagcgctt ccgaaaatgc
     4441 aacgcgagct gcgcacatac agctcactgt tcacgtcgca cctatatctg cgtgttgcct
     4501 gtatatatat atacatgaga agaacggcat agtgcgtgtt tatgcttaaa tgcgtactta
     4561 tatgcgtcta tttatgtagg atgaaaggta gtctagtacc tcctgtgata ttatcccatt
     4621 ccatgcgggg tatcgtatgc ttccttcagc actacccttt agctgttcta tatgctgcca
     4681 ctcctcaatt ggattagtct catccttcaa tgctatcatt tcctttgata ttggatcgat
     4741 ccgatgataa gctgtcaaac atgagaattg ggtaataact gatataatta aattgaagct
     4801 ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta gttttgctgg
     4861 ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta
     4921 gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag
     4981 accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc
     5041 acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga
     5101 gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta
     5161 tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta
     5221 ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc
     5281 acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac
     5341 tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg
     5401 tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag
     5461 atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt
     5521 aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg
     5581 atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta
     5641 gctttcgaca tgatttatct tcgtttcctg catgtttttg ttctgtgcag ttgggttaag
     5701 aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt
     5761 ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa
     5821 ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag ctaattcttg
     5881 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5941 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6001 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6061 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6121 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6181 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6241 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6301 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6361 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6421 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6481 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6541 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6601 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6661 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6721 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6781 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6841 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6901 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6961 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7021 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7081 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7141 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7201 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7261 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7321 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7381 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7441 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7501 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7561 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7621 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7681 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7741 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7801 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     7861 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     7921 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     7981 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     8041 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     8101 tgaccatgat tacgccaagc ttaccgcatc aggaaattgt aagcgttaat attttgttaa
     8161 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     8221 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     8281 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     8341 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     8401 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     8461 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     8521 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     8581 agggcgcgtc cattcgc
//
