LOCUS       sequence-332005-        8421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET fusion between ymTq2 and AausFP1 for use in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8421)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 8421)
  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-AausFP1 pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..8421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    197..221
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1708..1732)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1794..1813)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1795..1813)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1961..2148
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2016..2040)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2160..2177)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2160..2176)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2169..2191)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2612..3958
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4069..4872)
                     /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(4873..5088)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5113..5131)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5199..5303
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5304..5372,5373..6164)
                     /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(5522..5541)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6335..6923
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6824..6843
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7077..7094
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7211..7232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7247..7252,7253..7270,7271..7277)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7285..7301
                     /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     7290..7312
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7309..7325
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7309..7325
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7355..7810)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7492..7513)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7704..7723
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caatagtgaa aatctttttt tcttctatat ctacaaaaac tttttttttc tatcaacctc
       61 gttgataaat tttttcttta acaatcgtta ataattaatt aattggaaaa taaccatttt
      121 ttctctcttt tatacacaca ttcaaaagaa agaaaaaaaa tataccccag cctcgactag
      181 tgaattcgcg gccatcacaa gtttgtacaa aaaagcaggc tgctagcact agtaagcttt
      241 taattaaatg gttagtaaag gtgaagaatt gttcactggt gtagtgccaa ttttagtaga
      301 attagatggt gacgttaacg gtcataagtt ctctgtaagc ggtgaaggtg aaggtgacgc
      361 aacatacggt aaattaactt tgaaattcat ttgtactaca ggtaaattac cagttccatg
      421 gccaactttg gttactactt tgtcttgggg tgttcaatgt ttcgctagat acccagatca
      481 tatgaagcaa catgatttct ttaagtctgc tatgccagaa ggttacgttc aagaaagaac
      541 tatcttcttc aaggatgatg gtaactataa gactagagct gaagttaagt tcgaaggaga
      601 tactttagtt aatagaattg aattgaaggg tattgatttt aaggaagatg gtaatatttt
      661 gggtcataag ttggaatata attacttctc tgataacgtt tacattactg ctgataagca
      721 aaagaacggt attaaggcga acttcaagat tagacataac attgaagatg gtggtgttca
      781 attagctgat cattaccaac aaaacactcc aattggtgac ggtccagttt tgttgccaga
      841 taaccattac ttgtctactc aatctaagtt gtctaaggat ccaaacgaaa agagagatca
      901 tatggttttg ttggaattcg ttactgctgc tgggatcact ttaggtatgg atgaattgta
      961 taaatccgga ctcagatctt cggatccacc ggtcgccacc atgagttacg gagcactttt
     1021 gttcagagaa aaaatcccct atgttgttga gatggaaggt gatgttgagg gaatgaagtt
     1081 ttcagtaaga ggtaaaggac atggtgatgc taacactggt aaaattgaag ccagtttcat
     1141 ctgcacaact ggtgaactac cagtaccttg gtcaagcatt ctaacaactg tcacatacgg
     1201 tgcacagtgt tttgcaaaat atccaaatga tatcaaagac tacccaaaaa gtgctatgcc
     1261 agaaggttac gttcaagagc gaacaatcac ttttgaaaac gacggcgttt ataagacacg
     1321 cgctgaagtc acgtacgaga aaggatcagt ttataaccga gttacgttga acggttcagg
     1381 atttaaaaaa ggcggtaaca tcttgggcaa aaaattggaa tttaactaca accctcattg
     1441 catctatgtt cttccagatg tacaaaataa tggaatcaaa tgctacatca acattgttca
     1501 tgacgtaatt ggaggaggac aaataattgc tgcgcaccaa caattaaata ctcctcttgg
     1561 cggtggacca gtcgacattc cacactacca tcatatacaa gcgcacacaa tactttctaa
     1621 agatcccaaa gaaacaagag atcatatgaa cgtagtagag gtgttcagag ctatcgattg
     1681 caaaaccgca tatgcataag cggccgcacc cagctttctt gtacaaagtg gtgtagttaa
     1741 ttcatgatgg ccgctgcagg tcgacctcga gggggggccc ggtacccaat tcgccctata
     1801 gtgagtcgta ttacgcgcgg atccagcttt ggacttcttc gccagaggtt tggtcaagtc
     1861 tccaatcaag gttgtcggct tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg
     1921 tcaaatcgtt ggtagatacg ttgttgacac ttctaaataa gcgaatttct tatgatttat
     1981 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     2041 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     2101 ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggca tgccaattca
     2161 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     2221 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     2281 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
     2341 acgcatctgt gcggtatttc acaccgcata atcggatcgt acttgttacc catcattgaa
     2401 ttttgaacat ccgaacctgg gagttttccc tgaaacagat agtatatttg aacctgtata
     2461 ataatatata gtctagcgct ttacggaaga caatgtatgt atttcggttc ctggagaaac
     2521 tattgcatct attgcatagg taatcttgca cgtcgcatcc ccggttcatt ttctgcgttt
     2581 ccatcttgca cttcaatagc atatctttgt taacgaagca tctgtgcttc attttgtaga
     2641 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     2701 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     2761 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     2821 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     2881 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     2941 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     3001 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     3061 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     3121 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     3181 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     3241 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     3301 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     3361 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     3421 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     3481 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     3541 tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     3601 aataggaact tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg
     3661 caacgcgagc tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc
     3721 tgtatatata tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt
     3781 atatgcgtct atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat
     3841 tccatgcggg gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc
     3901 actcctcaat tggattagtc tcatccttca atgctatcat ttcctttgat attggatcga
     3961 tccgatgata agctgtcaaa catgagaatt gggtaataac tgatataatt aaattgaagc
     4021 tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg
     4081 gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt
     4141 agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga
     4201 gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc
     4261 cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg
     4321 agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt
     4381 atattctcca gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct
     4441 aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac
     4501 cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta
     4561 ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt
     4621 gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa
     4681 gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag
     4741 taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat
     4801 gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt
     4861 agctttcgac atgatttatc ttcgtttcct gcatgttttt gttctgtgca gttgggttaa
     4921 gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag
     4981 tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca
     5041 aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa gctaattctt
     5101 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5161 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5221 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5281 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5341 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5401 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5461 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5521 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5581 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5641 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5701 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5761 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5821 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5881 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5941 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6001 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6061 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6121 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6181 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6241 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6301 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6361 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6421 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6481 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6541 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6601 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6661 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6721 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6781 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6841 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6901 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6961 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7021 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     7081 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     7141 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     7201 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     7261 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     7321 atgaccatga ttacgccaag cttaccgcat caggaaattg taagcgttaa tattttgtta
     7381 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     7441 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     7501 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     7561 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     7621 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     7681 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     7741 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     7801 cagggcgcgt ccattcgcca agcttcctga aacggagaaa cataaacagg cattgctggg
     7861 atcacccata catcactctg ttttgcctga ccttttccgg taatttgaaa acaaacccgg
     7921 tctcgaagcg gagatccggc gataattacc gcagaaataa acccatacac gagacgtaga
     7981 accagccgca catggccgga gaaactcctg cgagaatttc gtaaactcgc gcgcattgca
     8041 tctgtatttc ctaatgcggc acttccaggc ctcgatcgag accgtttatc cattgctttt
     8101 ttgttgtctt tttccctcgt tcacagaaag tctgaagaag ctatagtaga actatgagct
     8161 ttttttgttt ctgttttcct tttttttttt tttacctctg tggaaattgt tactctcaca
     8221 ctctttagtt cgtttgtttg ttttgtttat tccaattatg accggtgacg aaacgtggtc
     8281 gatggtgggt accgcttatg ctcccctcca ttagtttcga ttatataaaa aggccaaata
     8341 ttgtattatt ttcaaatgtc ctatcattat cgtctaacat ctaatttctc ttaaattttt
     8401 tctctttctt tcctataaca c
//
