LOCUS       sequence-332108-        8433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET fusion between ymTq2 and ymNeongreen for use in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8433)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 8433)
  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-ymNeongreen pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..8433
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    88..109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(124..129,130..147,148..154)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    162..178
                     /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     167..189
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     186..202
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     186..202
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(232..687)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(369..390)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     581..600
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1495..1519
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2293..2302
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2554..2563
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3018..3042)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3104..3123)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3105..3123)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3271..3458
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3326..3350)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3470..3487)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3470..3486)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3479..3501)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3922..5268
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(5379..6182)
                     /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(6183..6398)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6423..6441)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6509..6613
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6614..6682,6683..7474)
                     /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(6832..6851)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7645..8233
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8134..8153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8387..8404
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
       61 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      121 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      181 tcacacagga aacagctatg accatgatta cgccaagctt accgcatcag gaaattgtaa
      241 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
      301 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
      361 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
      421 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
      481 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
      541 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
      601 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
      661 cgcttaatgc gccgctacag ggcgcgtcca ttcgccaagc ttcctgaaac ggagaaacat
      721 aaacaggcat tgctgggatc acccatacat cactctgttt tgcctgacct tttccggtaa
      781 tttgaaaaca aacccggtct cgaagcggag atccggcgat aattaccgca gaaataaacc
      841 catacacgag acgtagaacc agccgcacat ggccggagaa actcctgcga gaatttcgta
      901 aactcgcgcg cattgcatct gtatttccta atgcggcact tccaggcctc gatcgagacc
      961 gtttatccat tgcttttttg ttgtcttttt ccctcgttca cagaaagtct gaagaagcta
     1021 tagtagaact atgagctttt tttgtttctg ttttcctttt tttttttttt acctctgtgg
     1081 aaattgttac tctcacactc tttagttcgt ttgtttgttt tgtttattcc aattatgacc
     1141 ggtgacgaaa cgtggtcgat ggtgggtacc gcttatgctc ccctccatta gtttcgatta
     1201 tataaaaagg ccaaatattg tattattttc aaatgtccta tcattatcgt ctaacatcta
     1261 atttctctta aattttttct ctttctttcc tataacacca atagtgaaaa tctttttttc
     1321 ttctatatct acaaaaactt tttttttcta tcaacctcgt tgataaattt tttctttaac
     1381 aatcgttaat aattaattaa ttggaaaata accatttttt ctctctttta tacacacatt
     1441 caaaagaaag aaaaaaaata taccccagcc tcgactagtg aattcgcggc catcacaagt
     1501 ttgtacaaaa aagcaggctg ctagcactag taagctttta attaaatggt tagtaaaggt
     1561 gaagaattgt tcactggtgt agtgccaatt ttagtagaat tagatggtga cgttaacggt
     1621 cataagttct ctgtaagcgg tgaaggtgaa ggtgacgcaa catacggtaa attaactttg
     1681 aaattcattt gtactacagg taaattacca gttccatggc caactttggt tactactttg
     1741 tcttggggtg ttcaatgttt cgctagatac ccagatcata tgaagcaaca tgatttcttt
     1801 aagtctgcta tgccagaagg ttacgttcaa gaaagaacta tcttcttcaa ggatgatggt
     1861 aactataaga ctagagctga agttaagttc gaaggagata ctttagttaa tagaattgaa
     1921 ttgaagggta ttgattttaa ggaagatggt aatattttgg gtcataagtt ggaatataat
     1981 tacttctctg ataacgttta cattactgct gataagcaaa agaacggtat taaggcgaac
     2041 ttcaagatta gacataacat tgaagatggt ggtgttcaat tagctgatca ttaccaacaa
     2101 aacactccaa ttggtgacgg tccagttttg ttgccagata accattactt gtctactcaa
     2161 tctaagttgt ctaaggatcc aaacgaaaag agagatcata tggttttgtt ggaattcgtt
     2221 actgctgctg ggatcacttt aggtatggat gaattgtata aatccggact cagatcttcg
     2281 gatccaccgg tcgccaccat ggtctctaag ggtgaagaag ataacatggc ttctttgcca
     2341 gccacccacg agttgcacat cttcggttcc attaacggtg tcgacttcga tatggtcggt
     2401 caaggtaccg gtaacccaaa cgacggttac gaagaattga acttgaagtc taccaagggc
     2461 gacttgcagt tctccccttg gattttggtt ccacacatcg gttacggttt ccaccaatac
     2521 ttgccatacc cagacggtat gtccccattc caagccgcca tggttgacgg ttctggttac
     2581 caagtccaca gaaccatgca attcgaagac ggtgcttcct tgaccgtgaa ctacagatac
     2641 acctacgaag gttctcacat caagggcgaa gcccaagtta agggtaccgg tttccctgct
     2701 gacggtccag tcatgactaa ctccttgacc gctgctgact ggtgtcgttc caagaagacc
     2761 tacccaaacg ataagaccat catttccact ttcaagtggt cctacaccac cggtaacggc
     2821 aagagatacc gctctaccgc tcgtaccacc tacaccttcg ccaagccaat ggctgccaac
     2881 tacttgaaga accagcctat gtacgtcttc cgtaagaccg agctgaagca ttccaagact
     2941 gaattgaact tcaaggaatg gcaaaaggct ttcaccgacg tcatgggtat ggacgaattg
     3001 tacaagtaag cggccgcacc cagctttctt gtacaaagtg gtgtagttaa ttcatgatgg
     3061 ccgctgcagg tcgacctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta
     3121 ttacgcgcgg atccagcttt ggacttcttc gccagaggtt tggtcaagtc tccaatcaag
     3181 gttgtcggct tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt
     3241 ggtagatacg ttgttgacac ttctaaataa gcgaatttct tatgatttat gatttttatt
     3301 attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta
     3361 aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta
     3421 tagcatgagg tcgctcttat tgaccacacc tctaccggca tgccaattca ctggccgtcg
     3481 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     3541 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     3601 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     3661 gcggtatttc acaccgcata atcggatcgt acttgttacc catcattgaa ttttgaacat
     3721 ccgaacctgg gagttttccc tgaaacagat agtatatttg aacctgtata ataatatata
     3781 gtctagcgct ttacggaaga caatgtatgt atttcggttc ctggagaaac tattgcatct
     3841 attgcatagg taatcttgca cgtcgcatcc ccggttcatt ttctgcgttt ccatcttgca
     3901 cttcaatagc atatctttgt taacgaagca tctgtgcttc attttgtaga acaaaaatgc
     3961 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     4021 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     4081 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     4141 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     4201 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     4261 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     4321 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     4381 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     4441 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     4501 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     4561 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     4621 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     4681 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     4741 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     4801 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc
     4861 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact
     4921 tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc
     4981 tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata
     5041 tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct
     5101 atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg
     5161 gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat
     5221 tggattagtc tcatccttca atgctatcat ttcctttgat attggatcga tccgatgata
     5281 agctgtcaaa catgagaatt gggtaataac tgatataatt aaattgaagc tctaatttgt
     5341 gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt
     5401 ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt
     5461 ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca
     5521 tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt
     5581 gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag
     5641 ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca
     5701 gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt
     5761 gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt
     5821 gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg
     5881 actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg
     5941 gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca
     6001 tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg
     6061 gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat
     6121 agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac
     6181 atgatttatc ttcgtttcct gcatgttttt gttctgtgca gttgggttaa gaatactggg
     6241 caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc
     6301 ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga
     6361 aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa gctaattctt gaagacgaaa
     6421 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     6481 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6541 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6601 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6661 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6721 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6781 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6841 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6901 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     6961 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7021 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7081 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7141 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     7201 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     7261 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7321 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7381 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7441 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7501 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7561 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7621 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7681 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7741 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     7801 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7861 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7921 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7981 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     8041 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     8101 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     8161 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     8221 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     8281 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     8341 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     8401 cgaggaagcg gaagagcgcc caatacgcaa acc
//
