LOCUS       sequence-452205-        8632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TINGL (mTq2-based glucose sensor) in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8632)
  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 8632)
  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  pDRF1
FEATURES             Location/Qualifiers
     source          1..8632
                     /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(2522..2546)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2608..2627)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2609..2627)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2775..2962
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2830..2854)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2974..2991)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2974..2990)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2983..3005)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3426..4772
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4883..5686)
                     /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(5687..5902)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5927..5945)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6013..6117
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6118..6186,6187..6978)
                     /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(6336..6355)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7149..7737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7638..7657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7891..7908
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8025..8046
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8061..8066,8067..8084,8085..8091)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8099..8115
                     /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     8104..8126
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8123..8139
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8123..8139
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(8169..8624)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8306..8327)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8518..8537
                     /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 aagcttgagc
      841 tcatcgatat ggttagtaaa ggtgaagaat tgttcactgg tgtagtgcca attttagtag
      901 aattagatgg tgacgttaac ggtcataagt tctctgtaag cggtgaaggt gaaggtgacg
      961 caacatacgg taaattaact ttgaaattca tttgtactac aggtaaatta ccagttccat
     1021 ggccaacttt ggttactact ttgtcttggg gtgttcaatg tttcgctaga tacccagatc
     1081 atatgaagca acatgatttc tttaagtctg ctatgccaga aggttacgtt caagaaagaa
     1141 ctatcttctt caaggatgat ggtaactata agactagagc tgaagttaag ttcgaaggag
     1201 atactttagt taatagaatt gaattgaagg gtattgattt taaggaagat ggtaatattt
     1261 tgggtcataa gttggaatat aattacatgg ttgatactag cattggtgta acaatctata
     1321 agtacgacga ttattatatg tctgtagtgc gcaaggctat tgagcaagat gcgaaagccg
     1381 cgccagatgt tcagctgctg atgaatgatt ctcagaatga ccagtccaag cagaacgatc
     1441 agatcgacgt attgctggcg aaaggggtga aggcactggc aatcaacctg gttgacccgg
     1501 cagctgcggg tacggtgatt gagaaagcgc gtgggcaaaa cgtgccggtg gttttcttca
     1561 acaaagaacc gtctcgtaag gcgctggata gctacgacaa agcctactac gttggcactg
     1621 actccaaaga gtccggcatt attcaaggcg atttgattgc taaacactgg gcggcgaatc
     1681 agggttggga tctgaacaaa gacggtcaga ttcagttcgt actgctgaaa ggtgaaccgg
     1741 gccatccgga tgcagaagca cgtaccactt acgtgattaa agaattgaac gataaaggca
     1801 tcaaaactga acagttacag ttagataccg caatgtggga caccgctcag gcgaaagata
     1861 agatggacgc ctggctgtct ggcccgaacg ccaacaaaat cgaagtggtt atcgccaaca
     1921 acgatgcgat ggcaatgggc gcggttgaag cgctgaaagc acacaacaag tccagcattc
     1981 cggtgtttgg cgtcgatgcg ctgccagaag cgctggcgct ggtgaaatcc ggtgcattgg
     2041 cgggcaccgt actgaacgat gctaacaacc aggcgaaagc gacctttgat ctggcgaaaa
     2101 acctggccga tggtaaaggt gcggctgatg gcaccaactg gaaaatcgac aacaaagtgg
     2161 tccgcgtacc ttatgttggc gtagataaag acaacctggc tgagttcagc tacttctctg
     2221 ataacgttta cattactgct gataagcaaa agaacggtat taaggcgaac ttcaagatta
     2281 gacataacat tgaagatggt ggtgttcaat tagctgatca ttaccaacaa aacactccaa
     2341 ttggtgacgg tccagttttg ttgccagata accattactt gtctactcaa tctaagttgt
     2401 ctaaggatcc aaacgaaaag agagatcata tggttttgtt ggaattcgtt actgctgctg
     2461 ggatcacttt aggtatggat gaattgtata aataatgatc aagatctctg caggcggccg
     2521 cacccagctt tcttgtacaa agtggtgtag ttaattcatg atggccgctg caggtcgacc
     2581 tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacgc gcggatccag
     2641 ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc ggcttgtcta
     2701 ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga tacgttgttg
     2761 acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     2821 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
     2881 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat gaggtcgctc
     2941 ttattgacca cacctctacc ggcatgccaa ttcactggcc gtcgttttac aacgtcgtga
     3001 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     3061 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3121 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3181 cataatcgga tcgtacttgt tacccatcat tgaattttga acatccgaac ctgggagttt
     3241 tccctgaaac agatagtata tttgaacctg tataataata tatagtctag cgctttacgg
     3301 aagacaatgt atgtatttcg gttcctggag aaactattgc atctattgca taggtaatct
     3361 tgcacgtcgc atccccggtt cattttctgc gtttccatct tgcacttcaa tagcatatct
     3421 ttgttaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa
     3481 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc
     3541 tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc
     3601 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag
     3661 agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg
     3721 agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta
     3781 taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac
     3841 tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat
     3901 tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt atattctata
     3961 ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg
     4021 gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt
     4081 ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt
     4141 ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt
     4201 caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa
     4261 agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta
     4321 cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa
     4381 agcgctctga agttcctata ctttctagct agagaatagg aacttcggaa taggaacttc
     4441 aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca catacagctc
     4501 actgttcacg tcgcacctat atctgcgtgt tgcctgtata tatatataca tgagaagaac
     4561 ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc gtctatttat gtaggatgaa
     4621 aggtagtcta gtacctcctg tgatattatc ccattccatg cggggtatcg tatgcttcct
     4681 tcagcactac cctttagctg ttctatatgc tgccactcct caattggatt agtctcatcc
     4741 ttcaatgcta tcatttcctt tgatattgga tcgatccgat gataagctgt caaacatgag
     4801 aattgggtaa taactgatat aattaaattg aagctctaat ttgtgagttt agtatacatg
     4861 catttactta taatacagtt ttttagtttt gctggccgca tcttctcaaa tatgcttccc
     4921 agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt gcaaatagtc
     4981 ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg gttctatact
     5041 gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata atcaaccaat
     5101 cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata acaaaatctt
     5161 tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat agggagccct
     5221 tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact tcttctgccg
     5281 cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc gtaatgtctg
     5341 cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta ttaccaatgt
     5401 cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat gcctttagcg
     5461 gcttaactgt gccctccatg gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac
     5521 aaattttggg acctaatgct tcaactaact ccagtaattc cttggtggta cgaacatcca
     5581 atgaagcaca caagtttgtt tgcttttcgt gcatgatatt aaatagcttg gcagcaacag
     5641 gactaggatg agtagcagca cgttccttat atgtagcttt cgacatgatt tatcttcgtt
     5701 tcctgcatgt ttttgttctg tgcagttggg ttaagaatac tgggcaattt catgtttctt
     5761 caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt cgttcttcct
     5821 tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa aaaaaagaat
     5881 aaaaaaaaaa tgatgaattg aaaagctaat tcttgaagac gaaagggcct cgtgatacgc
     5941 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     6001 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6061 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6121 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6181 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6241 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6301 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6361 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6421 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6481 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6541 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6601 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6661 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6721 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6781 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6841 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6901 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6961 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7021 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7081 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7141 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7201 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7261 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7321 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7381 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7441 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7501 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7561 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7621 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7681 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7741 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7801 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7861 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7921 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7981 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     8041 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     8101 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttacc
     8161 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     8221 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     8281 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     8341 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     8401 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     8461 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     8521 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     8581 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gc
//
