LOCUS       sequence-236956-        8401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of yeast codon-optimized tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8401)
  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 8401)
  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 ytdTomato
FEATURES             Location/Qualifiers
     source          1..8401
                     /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(2291..2315)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2377..2396)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2378..2396)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2544..2731
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2599..2623)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2743..2760)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2743..2759)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2752..2774)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3195..4541
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4652..5455)
                     /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(5456..5671)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5696..5714)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5782..5886
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5887..5955,5956..6747)
                     /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(6105..6124)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6918..7506
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7407..7426
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7660..7677
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7794..7815
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7830..7835,7836..7853,7854..7860)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7868..7884
                     /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     7873..7895
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7892..7908
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7892..7908
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7938..8393)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8075..8096)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8287..8306
                     /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 ttttaaatta aatggttagt aaaggtgagg aagtgattaa ggaattcatg aggtttaaag
      901 ttagaatgga aggttctatg aatggtcatg aatttgaaat tgaaggtgaa ggtgaaggta
      961 gaccatatga aggtacacaa actgctaaat tgaaagttac aaaaggtggt ccattaccat
     1021 ttgcatggga tattttatct ccacagttta tgtatggtag taaagcatat gttaaacatc
     1081 cagctgatat tccagattat aagaaattaa gttttcctga aggttttaaa tgggaaagag
     1141 taatgaattt tgaagatggt ggtttagtaa cagttactca agattcttct ttacaagatg
     1201 gtacattaat atataaagtt aaaatgagag gtactaattt tccacctgat ggtccagtta
     1261 tgcagaagaa aacaatgggt tgggaagcct caactgaaag attgtaccct agagatggag
     1321 ttttgaaagg tgaaattcat caagctctta agttaaaaga tggaggtcac tatttggttg
     1381 agtttaaaac tatatacatg gctaaaaaac ctgttcaatt gccaggttat tattatgttg
     1441 atactaaatt agatattact tcacataatg aagattatac tattgttgaa caatatgaaa
     1501 gatctgaagg tagacatcat ttgtttttgg gtcatggtac tggtagtact ggttctggtt
     1561 cttcaggtac tgcttctagt gaagataata atatggctgt aatcaaagag ttcatgaggt
     1621 ttaaagttag aatggaaggt tctatgaatg gtcatgagtt tgaaatcgaa ggtgaaggag
     1681 aaggtagacc atatgaaggt actcaaactg ctaaattgaa agttactaaa ggtggtccat
     1741 tgccatttgc ttgggatata ttgtcacctc agtttatgta tggttctaaa gcatatgtta
     1801 aacatccagc tgatattcct gattataaaa aattatcatt tcctgaaggt tttaaatggg
     1861 aaagagttat gaattttgaa gatggtggtt tagttactgt aactcaagat tcttcattgc
     1921 aagatggtac attaatatat aaagttaaaa tgagaggtac taattttcca cctgatggtc
     1981 cagttatgca gaaaaagact atgggttggg aagcatctac tgaaagatta tatcctagag
     2041 atggtgtttt aaaaggtgaa attcaccaag ccttaaaatt gaaagatggt ggtcattatt
     2101 tggttgaatt caaaacaatc tacatggcta aaaaaccagt tcagctgccc ggatactatt
     2161 atgtcgatac taaactggat attacttcac acaatgaaga ctacacaatt gttgaacagt
     2221 atgaaagatc agagggtaga catcacttgt ttttatacgg tatggatgaa ttatataaat
     2281 aagcggccgc acccagcttt cttgtacaaa gtggtgtagt taattcatga tggccgctgc
     2341 aggtcgacct cgaggggggg cccggtaccc aattcgccct atagtgagtc gtattacgcg
     2401 cggatccagc tttggacttc ttcgccagag gtttggtcaa gtctccaatc aaggttgtcg
     2461 gcttgtctac cttgccagaa atttacgaaa agatggaaaa gggtcaaatc gttggtagat
     2521 acgttgttga cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat
     2581 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
     2641 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagcatg
     2701 aggtcgctct tattgaccac acctctaccg gcatgccaat tcactggccg tcgttttaca
     2761 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     2821 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     2881 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2941 ttcacaccgc ataatcggat cgtacttgtt acccatcatt gaattttgaa catccgaacc
     3001 tgggagtttt ccctgaaaca gatagtatat ttgaacctgt ataataatat atagtctagc
     3061 gctttacgga agacaatgta tgtatttcgg ttcctggaga aactattgca tctattgcat
     3121 aggtaatctt gcacgtcgca tccccggttc attttctgcg tttccatctt gcacttcaat
     3181 agcatatctt tgttaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga
     3241 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     3301 cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa
     3361 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     3421 caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa
     3481 tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg
     3541 tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa
     3601 gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg
     3661 tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta
     3721 tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat
     3781 tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat
     3841 aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa
     3901 tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag
     3961 atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat
     4021 atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt
     4081 agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg
     4141 gttttcaaaa gcgctctgaa gttcctatac tttctagcta gagaatagga acttcggaat
     4201 aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa atgcaacgcg agctgcgcac
     4261 atacagctca ctgttcacgt cgcacctata tctgcgtgtt gcctgtatat atatatacat
     4321 gagaagaacg gcatagtgcg tgtttatgct taaatgcgta cttatatgcg tctatttatg
     4381 taggatgaaa ggtagtctag tacctcctgt gatattatcc cattccatgc ggggtatcgt
     4441 atgcttcctt cagcactacc ctttagctgt tctatatgct gccactcctc aattggatta
     4501 gtctcatcct tcaatgctat catttccttt gatattggat cgatccgatg ataagctgtc
     4561 aaacatgaga attgggtaat aactgatata attaaattga agctctaatt tgtgagttta
     4621 gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat
     4681 atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg
     4741 caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca tcatccacgg
     4801 ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg ggtgtcataa
     4861 tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata aagccgataa
     4921 caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct ccagtagata
     4981 gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc tttgttactt
     5041 cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg tgtgcattcg
     5101 taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat ttgactgtat
     5161 taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg gcggataatg
     5221 cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc acatgtgttt
     5281 ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc ttggtggtac
     5341 gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta aatagcttgg
     5401 cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc gacatgattt
     5461 atcttcgttt cctgcatgtt tttgttctgt gcagttgggt taagaatact gggcaatttc
     5521 atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc tccttccttc
     5581 gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa
     5641 aaaaagaata aaaaaaaaat gatgaattga aaagctaatt cttgaagacg aaagggcctc
     5701 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5761 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5821 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5881 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5941 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6001 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6061 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6121 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6181 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6241 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6301 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6361 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6421 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6481 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6541 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6601 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6661 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6721 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6781 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6841 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6901 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6961 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     7021 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     7081 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7141 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7201 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7261 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7321 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7381 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7441 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7501 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7561 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     7621 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7681 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     7741 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7801 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     7861 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     7921 aagcttaccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     7981 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     8041 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     8101 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     8161 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     8221 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     8281 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     8341 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     8401 c
//
