LOCUS       sequence-331867-        8457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET fusion between ymTq2 and ystagRFP for use in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8457)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 8457)
  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-ystagRFP pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..8457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    191..215
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      989..998
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1738..1762)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1824..1843)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1825..1843)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1991..2178
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2046..2070)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2190..2207)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2190..2206)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2199..2221)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2642..3988
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4099..4902)
                     /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(4903..5118)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5143..5161)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5229..5333
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5334..5402,5403..6194)
                     /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(5552..5571)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6365..6953
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6854..6873
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7107..7124
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7241..7262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7277..7282,7283..7300,7301..7307)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7315..7331
                     /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     7320..7342
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7339..7355
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7339..7355
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7385..7840)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7522..7543)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7734..7753
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgaaaatctt tttttcttct atatctacaa aaactttttt tttctatcaa cctcgttgat
       61 aaattttttc tttaacaatc gttaataatt aattaattgg aaaataacca ttttttctct
      121 cttttataca cacattcaaa agaaagaaaa aaaatatacc ccagcctcga ctagtgaatt
      181 cgcggccatc acaagtttgt acaaaaaagc aggctgctag cactagtaag cttttaatta
      241 aatggttagt aaaggtgaag aattgttcac tggtgtagtg ccaattttag tagaattaga
      301 tggtgacgtt aacggtcata agttctctgt aagcggtgaa ggtgaaggtg acgcaacata
      361 cggtaaatta actttgaaat tcatttgtac tacaggtaaa ttaccagttc catggccaac
      421 tttggttact actttgtctt ggggtgttca atgtttcgct agatacccag atcatatgaa
      481 gcaacatgat ttctttaagt ctgctatgcc agaaggttac gttcaagaaa gaactatctt
      541 cttcaaggat gatggtaact ataagactag agctgaagtt aagttcgaag gagatacttt
      601 agttaataga attgaattga agggtattga ttttaaggaa gatggtaata ttttgggtca
      661 taagttggaa tataattact tctctgataa cgtttacatt actgctgata agcaaaagaa
      721 cggtattaag gcgaacttca agattagaca taacattgaa gatggtggtg ttcaattagc
      781 tgatcattac caacaaaaca ctccaattgg tgacggtcca gttttgttgc cagataacca
      841 ttacttgtct actcaatcta agttgtctaa ggatccaaac gaaaagagag atcatatggt
      901 tttgttggaa ttcgttactg ctgctgggat cactttaggt atggatgaat tgtataaatc
      961 cggactcaga tcttcggatc caccggtcgc caccatggta tctaaaggtg aagagttgat
     1021 aaaggaaaat atgcacatga aactctatat ggaaggcaca gtgaataacc atcattttaa
     1081 gtgtacttca gaaggagaag gtaagcctta tgaggggaca caaactatga gaattaaagt
     1141 tgttgaagga ggtccactac cattcgcttt tgacatcctg gcaacgagct tcatgtatgg
     1201 ctccagaaca ttcataaacc atacacaagg tattccagat ttcttcaaac aaagttttcc
     1261 agaaggcttt acttgggaga gagttactac ctacgaggac gggggtgttt taactgcgac
     1321 acaggataca tcattacaag atggctgttt gatctacaac gtaaaaatca gaggtgtcaa
     1381 ttttccatct aacggacctg ttatgcagaa aaagaccttg ggttgggagg ctaatactga
     1441 aatgctatac ccagcagatg gaggattgga aggtaggaca gtcatggcat tgaaacttgt
     1501 aggaggcggt cacctaattt gcaactttaa gacaacctac agaagcaaaa agcctgctaa
     1561 aaatcttaaa atgccagggg tgtactacgt ggatcataga cttgaacgta tcaaggaagc
     1621 tgacaaagag acttacgttg aacaacacga agtcgccgtc gccagatact gtgacttacc
     1681 ttcaaagtta ggtcataaac tgaatggtat ggatgaattg tataagtaag cggccgcacc
     1741 cagctttctt gtacaaagtg gtgtagttaa ttcatgatgg ccgctgcagg tcgacctcga
     1801 gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgg atccagcttt
     1861 ggacttcttc gccagaggtt tggtcaagtc tccaatcaag gttgtcggct tgtctacctt
     1921 gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt ggtagatacg ttgttgacac
     1981 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
     2041 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
     2101 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg tcgctcttat
     2161 tgaccacacc tctaccggca tgccaattca ctggccgtcg ttttacaacg tcgtgactgg
     2221 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     2281 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2341 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2401 atcggatcgt acttgttacc catcattgaa ttttgaacat ccgaacctgg gagttttccc
     2461 tgaaacagat agtatatttg aacctgtata ataatatata gtctagcgct ttacggaaga
     2521 caatgtatgt atttcggttc ctggagaaac tattgcatct attgcatagg taatcttgca
     2581 cgtcgcatcc ccggttcatt ttctgcgttt ccatcttgca cttcaatagc atatctttgt
     2641 taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
     2701 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
     2761 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
     2821 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
     2881 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
     2941 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
     3001 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
     3061 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
     3121 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
     3181 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
     3241 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
     3301 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
     3361 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
     3421 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
     3481 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
     3541 ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg
     3601 ctctgaagtt cctatacttt ctagctagag aataggaact tcggaatagg aacttcaaag
     3661 cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc tgcgcacata cagctcactg
     3721 ttcacgtcgc acctatatct gcgtgttgcc tgtatatata tatacatgag aagaacggca
     3781 tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct atttatgtag gatgaaaggt
     3841 agtctagtac ctcctgtgat attatcccat tccatgcggg gtatcgtatg cttccttcag
     3901 cactaccctt tagctgttct atatgctgcc actcctcaat tggattagtc tcatccttca
     3961 atgctatcat ttcctttgat attggatcga tccgatgata agctgtcaaa catgagaatt
     4021 gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta tacatgcatt
     4081 tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg cttcccagcc
     4141 tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa atagtcctct
     4201 tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc tatactgttg
     4261 acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca accaatcgta
     4321 accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa aatctttgtc
     4381 gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg agcccttgca
     4441 tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt ctgccgcctg
     4501 cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca
     4561 ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac caatgtcagc
     4621 aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct ttagcggctt
     4681 aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta gtaaacaaat
     4741 tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa catccaatga
     4801 agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag caacaggact
     4861 aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc ttcgtttcct
     4921 gcatgttttt gttctgtgca gttgggttaa gaatactggg caatttcatg tttcttcaac
     4981 actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt cttccttctg
     5041 ttcggagatt accgaatcaa aaaaatttca aggaaaccga aatcaaaaaa aagaataaaa
     5101 aaaaaatgat gaattgaaaa gctaattctt gaagacgaaa gggcctcgtg atacgcctat
     5161 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5221 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5281 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5341 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5401 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5461 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5521 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5581 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5641 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5701 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5761 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5821 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5881 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5941 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6001 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6061 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6121 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6181 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6241 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6301 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6361 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6421 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6481 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6541 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6601 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6661 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6721 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6781 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6841 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6901 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6961 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7021 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     7081 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     7141 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     7201 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     7261 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     7321 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cttaccgcat
     7381 caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     7441 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     7501 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     7561 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     7621 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     7681 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     7741 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     7801 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca agcttcctga
     7861 aacggagaaa cataaacagg cattgctggg atcacccata catcactctg ttttgcctga
     7921 ccttttccgg taatttgaaa acaaacccgg tctcgaagcg gagatccggc gataattacc
     7981 gcagaaataa acccatacac gagacgtaga accagccgca catggccgga gaaactcctg
     8041 cgagaatttc gtaaactcgc gcgcattgca tctgtatttc ctaatgcggc acttccaggc
     8101 ctcgatcgag accgtttatc cattgctttt ttgttgtctt tttccctcgt tcacagaaag
     8161 tctgaagaag ctatagtaga actatgagct ttttttgttt ctgttttcct tttttttttt
     8221 tttacctctg tggaaattgt tactctcaca ctctttagtt cgtttgtttg ttttgtttat
     8281 tccaattatg accggtgacg aaacgtggtc gatggtgggt accgcttatg ctcccctcca
     8341 ttagtttcga ttatataaaa aggccaaata ttgtattatt ttcaaatgtc ctatcattat
     8401 cgtctaacat ctaatttctc ttaaattttt tctctttctt tcctataaca ccaatag
//
