LOCUS       sequence-332106-        8439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET fusion between ymTq2 and mAvicFP1 for use in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8439)
  AUTHORS   Botman D, de Groot DH, Wehrens M, van Rossum K, Yska S, van der 
            Kieft B, Albert V, Minghetti N, Goedhart J, Teusink B
  TITLE     Side-by-side systematic characterization of novel FPs in budding 
            yeast
  JOURNAL   bioRxiv 2025.06.27.661975; doi: 
            https://doi.org/10.1101/2025.06.27.661975
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"; journalName: "bioRxiv 
            2025.06.27.661975; doi: https://doi.org/10.1101/2025.06.27.661975"
COMMENT     Sequence Label: ymTq2-mAvicFP1 pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..8439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(5..10,11..28,29..35)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    43..59
                     /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     48..70
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     67..83
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     67..83
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(113..568)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(250..271)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     462..481
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1376..1400
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2905..2929)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2991..3010)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2992..3010)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3158..3345
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3213..3237)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3357..3374)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3357..3373)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3366..3388)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3809..5155
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(5266..6069)
                     /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(6070..6285)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6310..6328)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6396..6500
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6501..6569,6570..7361)
                     /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(6719..6738)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7532..8120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8021..8040
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8274..8291
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8408..8429
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
       61 ttcacacagg aaacagctat gaccatgatt acgccaagct taccgcatca ggaaattgta
      121 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
      181 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
      241 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
      301 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
      361 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
      421 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
      481 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
      541 gcgcttaatg cgccgctaca gggcgcgtcc attcgccaag cttcctgaaa cggagaaaca
      601 taaacaggca ttgctgggat cacccataca tcactctgtt ttgcctgacc ttttccggta
      661 atttgaaaac aaacccggtc tcgaagcgga gatccggcga taattaccgc agaaataaac
      721 ccatacacga gacgtagaac cagccgcaca tggccggaga aactcctgcg agaatttcgt
      781 aaactcgcgc gcattgcatc tgtatttcct aatgcggcac ttccaggcct cgatcgagac
      841 cgtttatcca ttgctttttt gttgtctttt tccctcgttc acagaaagtc tgaagaagct
      901 atagtagaac tatgagcttt ttttgtttct gttttccttt tttttttttt tacctctgtg
      961 gaaattgtta ctctcacact ctttagttcg tttgtttgtt ttgtttattc caattatgac
     1021 cggtgacgaa acgtggtcga tggtgggtac cgcttatgct cccctccatt agtttcgatt
     1081 atataaaaag gccaaatatt gtattatttt caaatgtcct atcattatcg tctaacatct
     1141 aatttctctt aaattttttc tctttctttc ctataacacc aatagtgaaa atcttttttt
     1201 cttctatatc tacaaaaact ttttttttct atcaacctcg ttgataaatt ttttctttaa
     1261 caatcgttaa taattaatta attggaaaat aaccattttt tctctctttt atacacacat
     1321 tcaaaagaaa gaaaaaaaat ataccccagc ctcgactagt gaattcgcgg ccatcacaag
     1381 tttgtacaaa aaagcaggct gctagcacta gtaagctttt aattaaatgg ttagtaaagg
     1441 tgaagaattg ttcactggtg tagtgccaat tttagtagaa ttagatggtg acgttaacgg
     1501 tcataagttc tctgtaagcg gtgaaggtga aggtgacgca acatacggta aattaacttt
     1561 gaaattcatt tgtactacag gtaaattacc agttccatgg ccaactttgg ttactacttt
     1621 gtcttggggt gttcaatgtt tcgctagata cccagatcat atgaagcaac atgatttctt
     1681 taagtctgct atgccagaag gttacgttca agaaagaact atcttcttca aggatgatgg
     1741 taactataag actagagctg aagttaagtt cgaaggagat actttagtta atagaattga
     1801 attgaagggt attgatttta aggaagatgg taatattttg ggtcataagt tggaatataa
     1861 ttacttctct gataacgttt acattactgc tgataagcaa aagaacggta ttaaggcgaa
     1921 cttcaagatt agacataaca ttgaagatgg tggtgttcaa ttagctgatc attaccaaca
     1981 aaacactcca attggtgacg gtccagtttt gttgccagat aaccattact tgtctactca
     2041 atctaagttg tctaaggatc caaacgaaaa gagagatcat atggttttgt tggaattcgt
     2101 tactgctgct gggatcactt taggtatgga tgaattgtat aaatccggac tcagatcttc
     2161 ggatccaccg gtcgccacca tgtctaaggg tgctgaactg ttcaacggta tcgttcctat
     2221 cctgatcgaa ctgaacggtg acgttcatgg tcataagttc tctgttcgtg gtgaaggtga
     2281 aggtgacgct ggttctggta agatcgaaat caagttcgtt tgcaccaccg gtaccctgcc
     2341 tgttccttgg cctaccctgg ttaccacctt atgctacggt gttcagtgct tcacccgtta
     2401 ccctgaacat atgaagcagc atgacttcta caagtctgct atgcctgacg gttacatcca
     2461 gaagcgtacc atctctttcc aggacgacgg tcattacaag acccgtgctg aagttaagtt
     2521 cgaaggtgac accctggtta accgtatcga actgaagggt atcgacttca aggaagacgg
     2581 taacatcctg ggtaacaaga tggaatacaa ctacaactct cattctgttt acgttctgtc
     2641 tgacaaggct aacaacggta tcaaggttaa cttcaagatc cgtcataacc tgaagggtga
     2701 aggtatccag ctggctgacc atgaccagca gaacatccct atcggtgacg gtcctgttct
     2761 gctgcctgac taccattacc tgtctaccca gaccaagatc accaaggacc ctaacgaaaa
     2821 gcgtgaccat atgaacctgg ttgaattcgt taccgcttgc ggtatcaccc atggtatgga
     2881 cgaactgtac aagtaagcgg ccgcacccag ctttcttgta caaagtggtg tagttaattc
     2941 atgatggccg ctgcaggtcg acctcgaggg ggggcccggt acccaattcg ccctatagtg
     3001 agtcgtatta cgcgcggatc cagctttgga cttcttcgcc agaggtttgg tcaagtctcc
     3061 aatcaaggtt gtcggcttgt ctaccttgcc agaaatttac gaaaagatgg aaaagggtca
     3121 aatcgttggt agatacgttg ttgacacttc taaataagcg aatttcttat gatttatgat
     3181 ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta
     3241 ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc
     3301 tcaggtatag catgaggtcg ctcttattga ccacacctct accggcatgc caattcactg
     3361 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     3421 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     3481 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     3541 catctgtgcg gtatttcaca ccgcataatc ggatcgtact tgttacccat cattgaattt
     3601 tgaacatccg aacctgggag ttttccctga aacagatagt atatttgaac ctgtataata
     3661 atatatagtc tagcgcttta cggaagacaa tgtatgtatt tcggttcctg gagaaactat
     3721 tgcatctatt gcataggtaa tcttgcacgt cgcatccccg gttcattttc tgcgtttcca
     3781 tcttgcactt caatagcata tctttgttaa cgaagcatct gtgcttcatt ttgtagaaca
     3841 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     3901 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     3961 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     4021 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     4081 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
     4141 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     4201 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     4261 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     4321 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     4381 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     4441 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     4501 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     4561 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     4621 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     4681 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtct
     4741 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     4801 aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa
     4861 cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt
     4921 atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg cgtacttata
     4981 tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt atcccattcc
     5041 atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata tgctgccact
     5101 cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt ggatcgatcc
     5161 gatgataagc tgtcaaacat gagaattggg taataactga tataattaaa ttgaagctct
     5221 aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt tttgctggcc
     5281 gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct ctaccttagc
     5341 atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc ctgtagagac
     5401 cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat ctaaacccac
     5461 accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt ctctttgagc
     5521 aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac ccttagtata
     5581 ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa ggcctctagg
     5641 ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg ggcccaccac
     5701 accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg cagagtactg
     5761 caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta
     5821 cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat cagtcaagat
     5881 atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta actccagtaa
     5941 ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt cgtgcatgat
     6001 attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct tatatgtagc
     6061 tttcgacatg atttatcttc gtttcctgca tgtttttgtt ctgtgcagtt gggttaagaa
     6121 tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca atctaagtct
     6181 gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa aatttcaagg
     6241 aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa ttgaaaagct aattcttgaa
     6301 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     6361 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6421 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6481 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6541 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6601 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6661 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6721 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6781 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6841 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6901 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6961 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     7021 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     7081 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     7141 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     7201 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     7261 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7321 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     7381 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7441 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7501 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7561 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7621 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     7681 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7741 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7801 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7861 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7921 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7981 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     8041 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     8101 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     8161 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     8221 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     8281 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     8341 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     8401 acgcaattaa tgtgagttag ctcactcatt aggcacccc
//
