LOCUS       sequence-332107-        8457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET fusion between ymTq2 and yotagRFPT 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  TITLE     Teusink unpublished plasmids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-yotagRFPT pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..8457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    87..108
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(123..128,129..146,147..153)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    161..177
                     /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     166..188
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     185..201
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     185..201
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(231..686)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(368..389)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     580..599
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1494..1518
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2292..2301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3041..3065)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3127..3146)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3128..3146)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3294..3481
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3349..3373)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3493..3510)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3493..3509)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3502..3524)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3945..5291
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(5402..6205)
                     /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(6206..6421)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6446..6464)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6532..6636
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6637..6705,6706..7497)
                     /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(6855..6874)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7668..8256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8157..8176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8410..8427
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
       61 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
      121 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
      181 cacacaggaa acagctatga ccatgattac gccaagctta ccgcatcagg aaattgtaag
      241 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
      301 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
      361 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
      421 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
      481 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
      541 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
      601 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
      661 gcttaatgcg ccgctacagg gcgcgtccat tcgccaagct tcctgaaacg gagaaacata
      721 aacaggcatt gctgggatca cccatacatc actctgtttt gcctgacctt ttccggtaat
      781 ttgaaaacaa acccggtctc gaagcggaga tccggcgata attaccgcag aaataaaccc
      841 atacacgaga cgtagaacca gccgcacatg gccggagaaa ctcctgcgag aatttcgtaa
      901 actcgcgcgc attgcatctg tatttcctaa tgcggcactt ccaggcctcg atcgagaccg
      961 tttatccatt gcttttttgt tgtctttttc cctcgttcac agaaagtctg aagaagctat
     1021 agtagaacta tgagcttttt ttgtttctgt tttccttttt ttttttttta cctctgtgga
     1081 aattgttact ctcacactct ttagttcgtt tgtttgtttt gtttattcca attatgaccg
     1141 gtgacgaaac gtggtcgatg gtgggtaccg cttatgctcc cctccattag tttcgattat
     1201 ataaaaaggc caaatattgt attattttca aatgtcctat cattatcgtc taacatctaa
     1261 tttctcttaa attttttctc tttctttcct ataacaccaa tagtgaaaat ctttttttct
     1321 tctatatcta caaaaacttt ttttttctat caacctcgtt gataaatttt ttctttaaca
     1381 atcgttaata attaattaat tggaaaataa ccattttttc tctcttttat acacacattc
     1441 aaaagaaaga aaaaaaatat accccagcct cgactagtga attcgcggcc atcacaagtt
     1501 tgtacaaaaa agcaggctgc tagcactagt aagcttttaa ttaaatggtt agtaaaggtg
     1561 aagaattgtt cactggtgta gtgccaattt tagtagaatt agatggtgac gttaacggtc
     1621 ataagttctc tgtaagcggt gaaggtgaag gtgacgcaac atacggtaaa ttaactttga
     1681 aattcatttg tactacaggt aaattaccag ttccatggcc aactttggtt actactttgt
     1741 cttggggtgt tcaatgtttc gctagatacc cagatcatat gaagcaacat gatttcttta
     1801 agtctgctat gccagaaggt tacgttcaag aaagaactat cttcttcaag gatgatggta
     1861 actataagac tagagctgaa gttaagttcg aaggagatac tttagttaat agaattgaat
     1921 tgaagggtat tgattttaag gaagatggta atattttggg tcataagttg gaatataatt
     1981 acttctctga taacgtttac attactgctg ataagcaaaa gaacggtatt aaggcgaact
     2041 tcaagattag acataacatt gaagatggtg gtgttcaatt agctgatcat taccaacaaa
     2101 acactccaat tggtgacggt ccagttttgt tgccagataa ccattacttg tctactcaat
     2161 ctaagttgtc taaggatcca aacgaaaaga gagatcatat ggttttgttg gaattcgtta
     2221 ctgctgctgg gatcacttta ggtatggatg aattgtataa atccggactc agatcttcgg
     2281 atccaccggt cgccaccatg gtatctaaag gtgaagagtt gataaaggaa aatatgcaca
     2341 tgaaactcta tatggaaggc acagtgaata accatcattt taagtgtact tcagaaggag
     2401 aaggtaagcc ttatgagggg acacaaacta tgagaattaa agttgttgaa ggaggtccac
     2461 taccattcgc ttttgacatc ctggcaacga gcttcatgta tggctccaga acattcataa
     2521 accatacaca aggtattcca gatttcttca aacaaagttt tccagaaggc tttacttggg
     2581 agagagttac tacctacgag gacgggggtg ttttaactgc gacacaggat acatcattac
     2641 aagatggctg tttgatctac aacgtaaaaa tcagaggtgt caattttcca tctaacggac
     2701 ctgttatgca gaaaaagacc ttgggttggg aggctaatac tgaaatgcta tacccagcag
     2761 atggaggatt ggaaggtagg acagatatgg cattgaaact tgtaggaggc ggtcacctaa
     2821 tttgcaactt taagacaacc tacagaagca aaaagcctgc taaaaatctt aaaatgccag
     2881 gggtgtacta cgtggatcat agacttgaac gtatcaagga agctgacaaa gagacttacg
     2941 ttgaacaaca cgaagtcgcc gtcgccagat actgtgactt accttcaaag ttaggtcata
     3001 aactgaatgg tatggatgaa ttgtataagt aagcggccgc acccagcttt cttgtacaaa
     3061 gtggtgtagt taattcatga tggccgctgc aggtcgacct cgaggggggg cccggtaccc
     3121 aattcgccct atagtgagtc gtattacgcg cggatccagc tttggacttc ttcgccagag
     3181 gtttggtcaa gtctccaatc aaggttgtcg gcttgtctac cttgccagaa atttacgaaa
     3241 agatggaaaa gggtcaaatc gttggtagat acgttgttga cacttctaaa taagcgaatt
     3301 tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat
     3361 tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt
     3421 aggtcaggtt gctttctcag gtatagcatg aggtcgctct tattgaccac acctctaccg
     3481 gcatgccaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     3541 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     3601 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     3661 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataatcggat cgtacttgtt
     3721 acccatcatt gaattttgaa catccgaacc tgggagtttt ccctgaaaca gatagtatat
     3781 ttgaacctgt ataataatat atagtctagc gctttacgga agacaatgta tgtatttcgg
     3841 ttcctggaga aactattgca tctattgcat aggtaatctt gcacgtcgca tccccggttc
     3901 attttctgcg tttccatctt gcacttcaat agcatatctt tgttaacgaa gcatctgtgc
     3961 ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     4021 gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct
     4081 gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     4141 tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag
     4201 aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa
     4261 agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact
     4321 ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc
     4381 cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc
     4441 attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact
     4501 ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt
     4561 tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc
     4621 gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat
     4681 aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg
     4741 gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt
     4801 ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt
     4861 ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac
     4921 tttctagcta gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc
     4981 gcttccgaaa atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata
     5041 tctgcgtgtt gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct
     5101 taaatgcgta cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt
     5161 gatattatcc cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt
     5221 tctatatgct gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt
     5281 gatattggat cgatccgatg ataagctgtc aaacatgaga attgggtaat aactgatata
     5341 attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt
     5401 tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt
     5461 caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt
     5521 cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat gcgtctccct
     5581 tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac
     5641 ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa
     5701 cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca
     5761 tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa
     5821 tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata
     5881 cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcttcga
     5941 agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg
     6001 aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt
     6061 caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt
     6121 gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac
     6181 gttccttata tgtagctttc gacatgattt atcttcgttt cctgcatgtt tttgttctgt
     6241 gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat
     6301 ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat
     6361 caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga
     6421 aaagctaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     6481 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     6541 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6601 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6661 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6721 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6781 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6841 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6901 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6961 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     7021 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7081 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7141 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7201 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7261 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7321 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7381 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7441 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7501 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7561 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7621 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7681 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7741 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7801 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7861 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7921 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7981 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     8041 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     8101 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     8161 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     8221 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     8281 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     8341 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     8401 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccg
//
