LOCUS       sequence-332142-        8439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET fusion between ymTq2 and yeVenus for use in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8439)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 8439)
  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-yeVenus pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..8439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             503..1219
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     protein_bind    complement(1228..1252)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1314..1333)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1315..1333)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1481..1668
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1536..1560)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1680..1697)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1680..1696)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1689..1711)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2132..3478
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3589..4392)
                     /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(4393..4608)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4633..4651)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4719..4823
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4824..4892,4893..5684)
                     /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(5042..5061)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5855..6443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6344..6363
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6597..6614
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6731..6752
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6767..6772,6773..6790,6791..6797)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6805..6821
                     /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     6810..6832
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6829..6845
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6829..6845
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(6875..7330)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7012..7033)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7224..7243
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    8138..8162
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ctttaagtct gctatgccag aaggttacgt tcaagaaaga actatcttct tcaaggatga
       61 tggtaactat aagactagag ctgaagttaa gttcgaagga gatactttag ttaatagaat
      121 tgaattgaag ggtattgatt ttaaggaaga tggtaatatt ttgggtcata agttggaata
      181 taattacttc tctgataacg tttacattac tgctgataag caaaagaacg gtattaaggc
      241 gaacttcaag attagacata acattgaaga tggtggtgtt caattagctg atcattacca
      301 acaaaacact ccaattggtg acggtccagt tttgttgcca gataaccatt acttgtctac
      361 tcaatctaag ttgtctaagg atccaaacga aaagagagat catatggttt tgttggaatt
      421 cgttactgct gctgggatca ctttaggtat ggatgaattg tataaatccg gactcagatc
      481 ttcggatcca ccggtcgcca ccatgtctaa aggtgaagaa ttattcactg gtgttgtccc
      541 aattttggtt gaattagatg gtgatgttaa tggtcacaaa ttttctgtct ccggtgaagg
      601 tgaaggtgat gctacttacg gtaaattgac cttaaaattg atttgtacta ctggtaaatt
      661 gccagttcca tggccaacct tagtcactac tttaggttat ggtttgcaat gttttgctag
      721 atacccagat catatgaaac aacatgactt tttcaagtct gccatgccag aaggttatgt
      781 tcaagaaaga actatttttt tcaaagatga cggtaactac aagaccagag ctgaagtcaa
      841 gtttgaaggt gataccttag ttaatagaat cgaattaaaa ggtattgatt ttaaagaaga
      901 tggtaacatt ttaggtcaca aattggaata caactataac tctcacaatg tttacatcac
      961 tgctgacaaa caaaagaatg gtatcaaagc taacttcaaa attagacaca acattgaaga
     1021 tggtggtgtt caattagctg accattatca acaaaatact ccaattggtg atggtccagt
     1081 cttgttacca gacaaccatt acttatccta tcaatctgcc ttatccaaag atccaaacga
     1141 aaagagagac cacatggtct tgttagaatt tgttactgct gctggtatta cccatggtat
     1201 ggatgaattg tacaaataag cggccgcacc cagctttctt gtacaaagtg gtgtagttaa
     1261 ttcatgatgg ccgctgcagg tcgacctcga gggggggccc ggtacccaat tcgccctata
     1321 gtgagtcgta ttacgcgcgg atccagcttt ggacttcttc gccagaggtt tggtcaagtc
     1381 tccaatcaag gttgtcggct tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg
     1441 tcaaatcgtt ggtagatacg ttgttgacac ttctaaataa gcgaatttct tatgatttat
     1501 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     1561 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     1621 ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggca tgccaattca
     1681 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     1741 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     1801 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
     1861 acgcatctgt gcggtatttc acaccgcata atcggatcgt acttgttacc catcattgaa
     1921 ttttgaacat ccgaacctgg gagttttccc tgaaacagat agtatatttg aacctgtata
     1981 ataatatata gtctagcgct ttacggaaga caatgtatgt atttcggttc ctggagaaac
     2041 tattgcatct attgcatagg taatcttgca cgtcgcatcc ccggttcatt ttctgcgttt
     2101 ccatcttgca cttcaatagc atatctttgt taacgaagca tctgtgcttc attttgtaga
     2161 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     2221 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     2281 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     2341 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     2401 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     2461 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     2521 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     2581 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     2641 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     2701 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     2761 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     2821 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     2881 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     2941 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     3001 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     3061 tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     3121 aataggaact tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg
     3181 caacgcgagc tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc
     3241 tgtatatata tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt
     3301 atatgcgtct atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat
     3361 tccatgcggg gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc
     3421 actcctcaat tggattagtc tcatccttca atgctatcat ttcctttgat attggatcga
     3481 tccgatgata agctgtcaaa catgagaatt gggtaataac tgatataatt aaattgaagc
     3541 tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg
     3601 gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt
     3661 agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga
     3721 gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc
     3781 cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg
     3841 agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt
     3901 atattctcca gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct
     3961 aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac
     4021 cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta
     4081 ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt
     4141 gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa
     4201 gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag
     4261 taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat
     4321 gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt
     4381 agctttcgac atgatttatc ttcgtttcct gcatgttttt gttctgtgca gttgggttaa
     4441 gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag
     4501 tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca
     4561 aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa gctaattctt
     4621 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4681 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4741 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4801 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4861 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4921 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4981 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5041 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5101 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5161 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5221 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5281 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5341 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5401 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5461 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5521 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5581 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5641 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5701 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5761 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5821 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5881 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5941 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6001 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6061 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6121 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6181 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6241 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6301 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6361 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6421 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6481 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6541 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6601 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     6661 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     6721 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     6781 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     6841 atgaccatga ttacgccaag cttaccgcat caggaaattg taagcgttaa tattttgtta
     6901 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     6961 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     7021 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     7081 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     7141 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     7201 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     7261 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     7321 cagggcgcgt ccattcgcca agcttcctga aacggagaaa cataaacagg cattgctggg
     7381 atcacccata catcactctg ttttgcctga ccttttccgg taatttgaaa acaaacccgg
     7441 tctcgaagcg gagatccggc gataattacc gcagaaataa acccatacac gagacgtaga
     7501 accagccgca catggccgga gaaactcctg cgagaatttc gtaaactcgc gcgcattgca
     7561 tctgtatttc ctaatgcggc acttccaggc ctcgatcgag accgtttatc cattgctttt
     7621 ttgttgtctt tttccctcgt tcacagaaag tctgaagaag ctatagtaga actatgagct
     7681 ttttttgttt ctgttttcct tttttttttt tttacctctg tggaaattgt tactctcaca
     7741 ctctttagtt cgtttgtttg ttttgtttat tccaattatg accggtgacg aaacgtggtc
     7801 gatggtgggt accgcttatg ctcccctcca ttagtttcga ttatataaaa aggccaaata
     7861 ttgtattatt ttcaaatgtc ctatcattat cgtctaacat ctaatttctc ttaaattttt
     7921 tctctttctt tcctataaca ccaatagtga aaatcttttt ttcttctata tctacaaaaa
     7981 cttttttttt ctatcaacct cgttgataaa ttttttcttt aacaatcgtt aataattaat
     8041 taattggaaa ataaccattt tttctctctt ttatacacac attcaaaaga aagaaaaaaa
     8101 atatacccca gcctcgacta gtgaattcgc ggccatcaca agtttgtaca aaaaagcagg
     8161 ctgctagcac tagtaagctt ttaattaaat ggttagtaaa ggtgaagaat tgttcactgg
     8221 tgtagtgcca attttagtag aattagatgg tgacgttaac ggtcataagt tctctgtaag
     8281 cggtgaaggt gaaggtgacg caacatacgg taaattaact ttgaaattca tttgtactac
     8341 aggtaaatta ccagttccat ggccaacttt ggttactact ttgtcttggg gtgttcaatg
     8401 tttcgctaga tacccagatc atatgaagca acatgattt
//
