LOCUS       sequence-452206-        9364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TINGL-RR (mTq2-based glucose sensor - O2A - ymScarletI) in
            yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9364)
  AUTHORS   Botman D, Tielman A, Goedhart J, Teusink B
  TITLE     An mTurquoise2-based glucose biosensor
  JOURNAL   bioRxiv 2024.11.29.626064
REFERENCE   2  (bases 1 to 9364)
  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 
            2024.11.29.626064"
COMMENT     Sequence Label: TINGL-RR  pDRF1
FEATURES             Location/Qualifiers
     source          1..9364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    800..824
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3254..3278)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3340..3359)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3341..3359)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3507..3694
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3562..3586)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3706..3723)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3706..3722)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3715..3737)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4158..5504
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(5615..6418)
                     /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(6419..6634)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6659..6677)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6745..6849
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6850..6918,6919..7710)
                     /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(7068..7087)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7881..8469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8370..8389
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8623..8640
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8757..8778
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8793..8798,8799..8816,8817..8823)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8831..8847
                     /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     8836..8858
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8855..8871
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8855..8871
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(8901..9356)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9038..9059)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9250..9269
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caagcttcct gaaacggaga aacataaaca ggcattgctg ggatcaccca tacatcactc
       61 tgttttgcct gaccttttcc ggtaatttga aaacaaaccc ggtctcgaag cggagatccg
      121 gcgataatta ccgcagaaat aaacccatac acgagacgta gaaccagccg cacatggccg
      181 gagaaactcc tgcgagaatt tcgtaaactc gcgcgcattg catctgtatt tcctaatgcg
      241 gcacttccag gcctcgatcg agaccgttta tccattgctt ttttgttgtc tttttccctc
      301 gttcacagaa agtctgaaga agctatagta gaactatgag ctttttttgt ttctgttttc
      361 cttttttttt tttttacctc tgtggaaatt gttactctca cactctttag ttcgtttgtt
      421 tgttttgttt attccaatta tgaccggtga cgaaacgtgg tcgatggtgg gtaccgctta
      481 tgctcccctc cattagtttc gattatataa aaaggccaaa tattgtatta ttttcaaatg
      541 tcctatcatt atcgtctaac atctaatttc tcttaaattt tttctctttc tttcctataa
      601 caccaatagt gaaaatcttt ttttcttcta tatctacaaa aacttttttt ttctatcaac
      661 ctcgttgata aattttttct ttaacaatcg ttaataatta attaattgga aaataaccat
      721 tttttctctc ttttatacac acattcaaaa gaaagaaaaa aaatataccc cagcctcgac
      781 tagtgaattc gcggccatca caagtttgta caaaaaagca ggctgctagc atggttagta
      841 aaggtgaaga attgttcact ggtgtagtgc caattttagt agaattagat ggtgacgtta
      901 acggtcataa gttctctgta agcggtgaag gtgaaggtga cgcaacatac ggtaaattaa
      961 ctttgaaatt catttgtact acaggtaaat taccagttcc atggccaact ttggttacta
     1021 ctttgtcttg gggtgttcaa tgtttcgcta gatacccaga tcatatgaag caacatgatt
     1081 tctttaagtc tgctatgcca gaaggttacg ttcaagaaag aactatcttc ttcaaggatg
     1141 atggtaacta taagactaga gctgaagtta agttcgaagg agatacttta gttaatagaa
     1201 ttgaattgaa gggtattgat tttaaggaag atggtaatat tttgggtcat aagttggaat
     1261 ataattacat ggttgatact agcattggtg taacaatcta taagtacgac gattattata
     1321 tgtctgtagt gcgcaaggct attgagcaag atgcgaaagc cgcgccagat gttcagctgc
     1381 tgatgaatga ttctcagaat gaccagtcca agcagaacga tcagatcgac gtattgctgg
     1441 cgaaaggggt gaaggcactg gcaatcaacc tggttgaccc ggcagctgcg ggtacggtga
     1501 ttgagaaagc gcgtgggcaa aacgtgccgg tggttttctt caacaaagaa ccgtctcgta
     1561 aggcgctgga tagctacgac aaagcctact acgttggcac tgactccaaa gagtccggca
     1621 ttattcaagg cgatttgatt gctaaacact gggcggcgaa tcagggttgg gatctgaaca
     1681 aagacggtca gattcagttc gtactgctga aaggtgaacc gggccatccg gatgcagaag
     1741 cacgtaccac ttacgtgatt aaagaattga acgataaagg catcaaaact gaacagttac
     1801 agttagatac cgcaatgtgg gacaccgctc aggcgaaaga taagatggac gcctggctgt
     1861 ctggcccgaa cgccaacaaa atcgaagtgg ttatcgccaa caacgatgcg atggcaatgg
     1921 gcgcggttga agcgctgaaa gcacacaaca agtccagcat tccggtgttt ggcgtcgatg
     1981 cgctgccaga agcgctggcg ctggtgaaat ccggtgcatt ggcgggcacc gtactgaacg
     2041 atgctaacaa ccaggcgaaa gcgacctttg atctggcgaa aaacctggcc gatggtaaag
     2101 gtgcggctga tggcaccaac tggaaaatcg acaacaaagt ggtccgcgta ccttatgttg
     2161 gcgtagataa agacaacctg gctgagttca gctacttctc tgataacgtt tacattactg
     2221 ctgataagca aaagaacggt attaaggcga acttcaagat tagacataac attgaagatg
     2281 gtggtgttca attagctgat cattaccaac aaaacactcc aattggtgac ggtccagttt
     2341 tgttgccaga taaccattac ttgtctactc aatctaagtt gtctaaggat ccaaacgaaa
     2401 agagagatca tatggttttg ttggaattcg ttactgctgc tgggatcact ttaggtatgg
     2461 atgaattgta taaaggttct ggtaagtcca attacgacct attgaagttg tgtggagatg
     2521 ttgaatccaa cccaggacct cccgggatgg ttagtaaagg tgaagctgtt ataaaagaat
     2581 tcatgaggtt taaagttcat atggaaggtt caatgaatgg tcatgaattt gaaattgaag
     2641 gtgaaggtga aggtagacca tatgaaggta cacaaactgc taaattgaaa gttactaaag
     2701 gtggtccatt accattttct tgggatattt tgtctccaca attcatgtat ggttctagag
     2761 cttttattaa gcatccagct gatattccag attattataa acaatctttt cctgaaggtt
     2821 ttaaatggga aagagttatg aatttcgaag atggtggtgc tgttactgtt actcaagata
     2881 cttctttgga agatggtact ttaatctata aagttaaatt gagaggtact aattttccac
     2941 cagatggtcc agttatgcaa aagaaaacta tgggttggga agcatctact gaaagattgt
     3001 atccagaaga tggtgttttg aaaggagaca ttaagatggc tttgagattg aaagatggtg
     3061 gtcggtacct ggccgacttc aagaccacct ataaagctaa aaaaccagtt caaatgccag
     3121 gtgcatataa tgttgataga aagttagata taacgtcgca taacgaggac tatactgtag
     3181 ttgaacaata tgaacgtagt gaaggtagac atagtaccgg aggaatggat gaattgtata
     3241 aataagcggc cgcacccagc tttcttgtac aaagtggtgt agttaattca tgatggccgc
     3301 tgcaggtcga cctcgagggg gggcccggta cccaattcgc cctatagtga gtcgtattac
     3361 gcgcggatcc agctttggac ttcttcgcca gaggtttggt caagtctcca atcaaggttg
     3421 tcggcttgtc taccttgcca gaaatttacg aaaagatgga aaagggtcaa atcgttggta
     3481 gatacgttgt tgacacttct aaataagcga atttcttatg atttatgatt tttattatta
     3541 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     3601 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagc
     3661 atgaggtcgc tcttattgac cacacctcta ccggcatgcc aattcactgg ccgtcgtttt
     3721 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     3781 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3841 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3901 tatttcacac cgcataatcg gatcgtactt gttacccatc attgaatttt gaacatccga
     3961 acctgggagt tttccctgaa acagatagta tatttgaacc tgtataataa tatatagtct
     4021 agcgctttac ggaagacaat gtatgtattt cggttcctgg agaaactatt gcatctattg
     4081 cataggtaat cttgcacgtc gcatccccgg ttcattttct gcgtttccat cttgcacttc
     4141 aatagcatat ctttgttaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg
     4201 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     4261 acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg
     4321 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     4381 atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa
     4441 aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct
     4501 ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta
     4561 gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc
     4621 gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga
     4681 ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat
     4741 gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg
     4801 tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta
     4861 caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt
     4921 tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga
     4981 gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt
     5041 agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt cagagcgctt
     5101 ttggttttca aaagcgctct gaagttccta tactttctag ctagagaata ggaacttcgg
     5161 aataggaact tcaaagcgtt tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg
     5221 cacatacagc tcactgttca cgtcgcacct atatctgcgt gttgcctgta tatatatata
     5281 catgagaaga acggcatagt gcgtgtttat gcttaaatgc gtacttatat gcgtctattt
     5341 atgtaggatg aaaggtagtc tagtacctcc tgtgatatta tcccattcca tgcggggtat
     5401 cgtatgcttc cttcagcact accctttagc tgttctatat gctgccactc ctcaattgga
     5461 ttagtctcat ccttcaatgc tatcatttcc tttgatattg gatcgatccg atgataagct
     5521 gtcaaacatg agaattgggt aataactgat ataattaaat tgaagctcta atttgtgagt
     5581 ttagtataca tgcatttact tataatacag ttttttagtt ttgctggccg catcttctca
     5641 aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca tcccttccct
     5701 ttgcaaatag tcctcttcca acaataataa tgtcagatcc tgtagagacc acatcatcca
     5761 cggttctata ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca
     5821 taatcaacca atcgtaacct tcatctcttc cacccatgtc tctttgagca ataaagccga
     5881 taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc cttagtatat tctccagtag
     5941 atagggagcc cttgcatgac aattctgcta acatcaaaag gcctctaggt tcctttgtta
     6001 cttcttctgc cgcctgcttc aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat
     6061 tcgtaatgtc tgcccattct gctattctgt atacacccgc agagtactgc aatttgactg
     6121 tattaccaat gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac ttggcggata
     6181 atgcctttag cggcttaact gtgccctcca tggaaaaatc agtcaagata tccacatgtg
     6241 tttttagtaa acaaattttg ggacctaatg cttcaactaa ctccagtaat tccttggtgg
     6301 tacgaacatc caatgaagca cacaagtttg tttgcttttc gtgcatgata ttaaatagct
     6361 tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct ttcgacatga
     6421 tttatcttcg tttcctgcat gtttttgttc tgtgcagttg ggttaagaat actgggcaat
     6481 ttcatgtttc ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc
     6541 ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc
     6601 aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaagcta attcttgaag acgaaagggc
     6661 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     6721 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6781 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6841 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6901 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6961 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     7021 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     7081 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     7141 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     7201 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     7261 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     7321 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7381 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7441 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7501 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7561 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7621 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7681 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7741 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7801 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7861 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7921 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7981 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     8041 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     8101 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     8161 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     8221 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     8281 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     8341 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8401 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8461 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8521 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     8581 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     8641 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     8701 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     8761 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     8821 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     8881 gccaagctta ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     8941 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     9001 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     9061 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     9121 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     9181 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     9241 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     9301 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     9361 tcgc
//
