LOCUS       sequence-452207-        9376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TINGL-RR (mTq2-based glucose sensor - O2A - ymNeongreen) 
            in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9376)
  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 9376)
  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-GR pDRF1
FEATURES             Location/Qualifiers
     source          1..9376
                     /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"
     regulatory      2802..2811
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3266..3290)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3352..3371)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3353..3371)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3519..3706
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3574..3598)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3718..3735)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3718..3734)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3727..3749)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4170..5516
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(5627..6430)
                     /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(6431..6646)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6671..6689)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6757..6861
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6862..6930,6931..7722)
                     /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(7080..7099)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7893..8481
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8382..8401
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8635..8652
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8769..8790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8805..8810,8811..8828,8829..8835)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8843..8859
                     /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     8848..8870
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8867..8883
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8867..8883
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(8913..9368)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9050..9071)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9262..9281
                     /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 tctctaaggg tgaagaagat aacatggctt
     2581 ctttgccagc cacccacgag ttgcacatct tcggttccat taacggtgtc gacttcgata
     2641 tggtcggtca aggtaccggt aacccaaacg acggttacga agaattgaac ttgaagtcta
     2701 ccaagggcga cttgcagttc tccccttgga ttttggttcc acacatcggt tacggtttcc
     2761 accaatactt gccataccca gacggtatgt ccccattcca agccgccatg gttgacggtt
     2821 ctggttacca agtccacaga accatgcaat tcgaagacgg tgcttccttg accgtgaact
     2881 acagatacac ctacgaaggt tctcacatca agggcgaagc ccaagttaag ggtaccggtt
     2941 tccctgctga cggtccagtc atgactaact ccttgaccgc tgctgactgg tgtcgttcca
     3001 agaagaccta cccaaacgat aagaccatca tttccacttt caagtggtcc tacaccaccg
     3061 gtaacggcaa gagataccgc tctaccgctc gtaccaccta caccttcgcc aagccaatgg
     3121 ctgccaacta cttgaagaac cagcctatgt acgtcttccg taagaccgag ctgaagcatt
     3181 ccaagactga attgaacttc aaggaatggc aaaaggcttt caccgacgtc atgggtatgg
     3241 acgaattata caagtaagcg gccgcaccca gctttcttgt acaaagtggt gtagttaatt
     3301 catgatggcc gctgcaggtc gacctcgagg gggggcccgg tacccaattc gccctatagt
     3361 gagtcgtatt acgcgcggat ccagctttgg acttcttcgc cagaggtttg gtcaagtctc
     3421 caatcaaggt tgtcggcttg tctaccttgc cagaaattta cgaaaagatg gaaaagggtc
     3481 aaatcgttgg tagatacgtt gttgacactt ctaaataagc gaatttctta tgatttatga
     3541 tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt
     3601 aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt
     3661 ctcaggtata gcatgaggtc gctcttattg accacacctc taccggcatg ccaattcact
     3721 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     3781 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     3841 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     3901 gcatctgtgc ggtatttcac accgcataat cggatcgtac ttgttaccca tcattgaatt
     3961 ttgaacatcc gaacctggga gttttccctg aaacagatag tatatttgaa cctgtataat
     4021 aatatatagt ctagcgcttt acggaagaca atgtatgtat ttcggttcct ggagaaacta
     4081 ttgcatctat tgcataggta atcttgcacg tcgcatcccc ggttcatttt ctgcgtttcc
     4141 atcttgcact tcaatagcat atctttgtta acgaagcatc tgtgcttcat tttgtagaac
     4201 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag
     4261 aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt
     4321 aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     4381 tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt
     4441 tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac
     4501 tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt
     4561 ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg
     4621 actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa
     4681 aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt
     4741 gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct
     4801 ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa
     4861 tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt
     4921 agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg
     4981 atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat
     5041 tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc
     5101 ttcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct agctagagaa
     5161 taggaacttc ggaataggaa cttcaaagcg tttccgaaaa cgagcgcttc cgaaaatgca
     5221 acgcgagctg cgcacataca gctcactgtt cacgtcgcac ctatatctgc gtgttgcctg
     5281 tatatatata tacatgagaa gaacggcata gtgcgtgttt atgcttaaat gcgtacttat
     5341 atgcgtctat ttatgtagga tgaaaggtag tctagtacct cctgtgatat tatcccattc
     5401 catgcggggt atcgtatgct tccttcagca ctacccttta gctgttctat atgctgccac
     5461 tcctcaattg gattagtctc atccttcaat gctatcattt cctttgatat tggatcgatc
     5521 cgatgataag ctgtcaaaca tgagaattgg gtaataactg atataattaa attgaagctc
     5581 taatttgtga gtttagtata catgcattta cttataatac agttttttag ttttgctggc
     5641 cgcatcttct caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag
     5701 catcccttcc ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga
     5761 ccacatcatc cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca
     5821 caccgggtgt cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag
     5881 caataaagcc gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat
     5941 attctccagt agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag
     6001 gttcctttgt tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca
     6061 caccgtgtgc attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact
     6121 gcaatttgac tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt
     6181 acttggcgga taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga
     6241 tatccacatg tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta
     6301 attccttggt ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga
     6361 tattaaatag cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag
     6421 ctttcgacat gatttatctt cgtttcctgc atgtttttgt tctgtgcagt tgggttaaga
     6481 atactgggca atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc
     6541 tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaag
     6601 gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaaaagc taattcttga
     6661 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     6721 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6781 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6841 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6901 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6961 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     7021 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     7081 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     7141 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     7201 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     7261 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     7321 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     7381 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     7441 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7501 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7561 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7621 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7681 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7741 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7801 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7861 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7921 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7981 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     8041 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     8101 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     8161 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     8221 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     8281 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     8341 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     8401 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     8461 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     8521 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     8581 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     8641 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     8701 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     8761 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     8821 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     8881 gaccatgatt acgccaagct taccgcatca ggaaattgta agcgttaata ttttgttaaa
     8941 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     9001 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     9061 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     9121 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     9181 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     9241 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     9301 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     9361 gggcgcgtcc attcgc
//
