LOCUS       sequence-332369-        8427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET fusion between ymTq2 and tdTomato for use in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8427)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 8427)
  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"
COMMENT     Sequence Label: ymTq2-tdTomato pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..8427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    196..220
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      994..1003
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1000..1701
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(1133..1151)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1401..1420
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1595..1618
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(1713..1737)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1799..1818)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1800..1818)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1966..2153
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2021..2045)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2165..2182)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2165..2181)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2174..2196)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2617..3963
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4074..4877)
                     /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(4878..5093)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5118..5136)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5204..5308
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5309..5377,5378..6169)
                     /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(5527..5546)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6340..6928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6829..6848
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7082..7099
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7216..7237
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7252..7257,7258..7275,7276..7282)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7290..7306
                     /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     7295..7317
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7314..7330
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7314..7330
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7360..7815)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7497..7518)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7709..7728
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 aatagtgaaa atcttttttt cttctatatc tacaaaaact ttttttttct atcaacctcg
       61 ttgataaatt ttttctttaa caatcgttaa taattaatta attggaaaat aaccattttt
      121 tctctctttt atacacacat tcaaaagaaa gaaaaaaaat ataccccagc ctcgactagt
      181 gaattcgcgg ccatcacaag tttgtacaaa aaagcaggct gctagcacta gtaagctttt
      241 aattaaatgg ttagtaaagg tgaagaattg ttcactggtg tagtgccaat tttagtagaa
      301 ttagatggtg acgttaacgg tcataagttc tctgtaagcg gtgaaggtga aggtgacgca
      361 acatacggta aattaacttt gaaattcatt tgtactacag gtaaattacc agttccatgg
      421 ccaactttgg ttactacttt gtcttggggt gttcaatgtt tcgctagata cccagatcat
      481 atgaagcaac atgatttctt taagtctgct atgccagaag gttacgttca agaaagaact
      541 atcttcttca aggatgatgg taactataag actagagctg aagttaagtt cgaaggagat
      601 actttagtta atagaattga attgaagggt attgatttta aggaagatgg taatattttg
      661 ggtcataagt tggaatataa ttacttctct gataacgttt acattactgc tgataagcaa
      721 aagaacggta ttaaggcgaa cttcaagatt agacataaca ttgaagatgg tggtgttcaa
      781 ttagctgatc attaccaaca aaacactcca attggtgacg gtccagtttt gttgccagat
      841 aaccattact tgtctactca atctaagttg tctaaggatc caaacgaaaa gagagatcat
      901 atggttttgt tggaattcgt tactgctgct gggatcactt taggtatgga tgaattgtat
      961 aaatccggac tcagatcttc ggatccaccg gtcgccacca tggtgagcaa gggcgaggag
     1021 gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag
     1081 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     1141 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg
     1201 tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc
     1261 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc
     1321 gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc
     1381 accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     1441 accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag
     1501 ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc
     1561 gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag
     1621 gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgtac
     1681 ggcatggacg agctgtacaa gtaagcggcc gcacccagct ttcttgtaca aagtggtgta
     1741 gttaattcat gatggccgct gcaggtcgac ctcgaggggg ggcccggtac ccaattcgcc
     1801 ctatagtgag tcgtattacg cgcggatcca gctttggact tcttcgccag aggtttggtc
     1861 aagtctccaa tcaaggttgt cggcttgtct accttgccag aaatttacga aaagatggaa
     1921 aagggtcaaa tcgttggtag atacgttgtt gacacttcta aataagcgaa tttcttatga
     1981 tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt
     2041 gactcttagg ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg
     2101 ttgctttctc aggtatagca tgaggtcgct cttattgacc acacctctac cggcatgcca
     2161 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     2221 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     2281 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
     2341 tccttacgca tctgtgcggt atttcacacc gcataatcgg atcgtacttg ttacccatca
     2401 ttgaattttg aacatccgaa cctgggagtt ttccctgaaa cagatagtat atttgaacct
     2461 gtataataat atatagtcta gcgctttacg gaagacaatg tatgtatttc ggttcctgga
     2521 gaaactattg catctattgc ataggtaatc ttgcacgtcg catccccggt tcattttctg
     2581 cgtttccatc ttgcacttca atagcatatc tttgttaacg aagcatctgt gcttcatttt
     2641 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     2701 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     2761 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     2821 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     2881 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     2941 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     3001 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     3061 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     3121 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     3181 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     3241 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     3301 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     3361 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     3421 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     3481 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     3541 gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc
     3601 tagagaatag gaacttcgga ataggaactt caaagcgttt ccgaaaacga gcgcttccga
     3661 aaatgcaacg cgagctgcgc acatacagct cactgttcac gtcgcaccta tatctgcgtg
     3721 ttgcctgtat atatatatac atgagaagaa cggcatagtg cgtgtttatg cttaaatgcg
     3781 tacttatatg cgtctattta tgtaggatga aaggtagtct agtacctcct gtgatattat
     3841 cccattccat gcggggtatc gtatgcttcc ttcagcacta ccctttagct gttctatatg
     3901 ctgccactcc tcaattggat tagtctcatc cttcaatgct atcatttcct ttgatattgg
     3961 atcgatccga tgataagctg tcaaacatga gaattgggta ataactgata taattaaatt
     4021 gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt
     4081 tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct
     4141 accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct
     4201 gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct
     4261 aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct
     4321 ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc
     4381 ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg
     4441 cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg
     4501 cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca
     4561 gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa
     4621 aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca
     4681 gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac
     4741 tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg
     4801 tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta
     4861 tatgtagctt tcgacatgat ttatcttcgt ttcctgcatg tttttgttct gtgcagttgg
     4921 gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat
     4981 ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa
     5041 tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaaaagctaa
     5101 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     5161 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5221 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5281 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5341 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5401 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5461 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5521 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5581 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5641 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5701 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5761 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5821 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5881 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5941 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6001 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6061 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6121 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6181 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6241 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6301 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6361 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6421 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6481 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6541 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6601 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6661 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6721 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6781 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6841 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6901 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6961 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     7021 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     7081 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     7141 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     7201 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     7261 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     7321 cagctatgac catgattacg ccaagcttac cgcatcagga aattgtaagc gttaatattt
     7381 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     7441 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     7501 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     7561 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     7621 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     7681 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     7741 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     7801 cgctacaggg cgcgtccatt cgccaagctt cctgaaacgg agaaacataa acaggcattg
     7861 ctgggatcac ccatacatca ctctgttttg cctgaccttt tccggtaatt tgaaaacaaa
     7921 cccggtctcg aagcggagat ccggcgataa ttaccgcaga aataaaccca tacacgagac
     7981 gtagaaccag ccgcacatgg ccggagaaac tcctgcgaga atttcgtaaa ctcgcgcgca
     8041 ttgcatctgt atttcctaat gcggcacttc caggcctcga tcgagaccgt ttatccattg
     8101 cttttttgtt gtctttttcc ctcgttcaca gaaagtctga agaagctata gtagaactat
     8161 gagctttttt tgtttctgtt ttcctttttt ttttttttac ctctgtggaa attgttactc
     8221 tcacactctt tagttcgttt gtttgttttg tttattccaa ttatgaccgg tgacgaaacg
     8281 tggtcgatgg tgggtaccgc ttatgctccc ctccattagt ttcgattata taaaaaggcc
     8341 aaatattgta ttattttcaa atgtcctatc attatcgtct aacatctaat ttctcttaaa
     8401 ttttttctct ttctttccta taacacc
//
