LOCUS       sequence-323180-        8276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression of the RNA-based sensor for 
            fructose-1,6-bisphosphate #4_1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8276)
  AUTHORS   Ortega AD, Takhaveev V, Vedelaar SR, Long Y, Mestre-Farras N, 
            Incarnato D, Ersoy F, Olsen LF, Mayer G, Heinemann M
  TITLE     A synthetic RNA-based biosensor for fructose-1,6-bisphosphate that 
            reports glycolytic flux.
  JOURNAL   Cell Chem Biol. 2021 Apr 22. pii: S2451-9456(21)00161-6. doi: 
            10.1016/j.chembiol.2021.04.006.
  PUBMED    33915105
REFERENCE   2  (bases 1 to 8276)
  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: "Cell Chem 
            Biol. 2021 Apr 22. pii: S2451-9456(21)00161-6. doi: 
            10.1016/j.chembiol.2021.04.006."
COMMENT     Sequence Label: pFBP-4_1.sensor
FEATURES             Location/Qualifiers
     source          1..8276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        194..409
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             410..1213
                     /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"
     rep_origin      complement(1344..1799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1481..1502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1693..1712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1925..1947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1939..1956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1940..1956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1963..1982
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1963..1981
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1995..2392
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      3148..3395
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3164..3182)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     3860..3879
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        3960..4356
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             4416..5132
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGIIHGMDELYK"
     terminator      5268..5455
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5323..5347)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        complement(5503..5521)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5542..5558)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5542..5558)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5555..5577)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5566..5582
                     /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)."
     promoter        complement(join(5590..5596,5597..5614,5615..5620))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5635..5656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5773..5790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5944..6532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6024..6043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6703..7494,7495..7563))
                     /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     7326..7345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7564..7668)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7705..8208
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8249..8267
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcggggg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
      361 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
      421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
      481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
      541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
      601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
      661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
      721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
      781 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
      841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
      901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
      961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1921 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag
     1981 ggcgaattgg agctcatagc ttcaaaatgt ttctactcct tttttactct tccagatttt
     2041 ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca agcacagcat actaaatttc
     2101 ccctctttct tcctctaggg tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa
     2161 agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt
     2221 ctttttcttg aaaatttttt tttgattttt ttctctttcg atgacctccc attgatattt
     2281 aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg ttctattaca
     2341 acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta agttttgccg
     2401 cgggaaataa tgtctatggt tagtaaagga gaagaaaata acatggcaat cattaaggag
     2461 ttcatgagat tcaaagttca catggaaggt tctgtaaatg gacatgaatt tgaaatagaa
     2521 ggtgaaggag aaggaaggcc ttatgaagga acccaaaccg cgaagctaaa agttactaag
     2581 ggtggcccat taccatttgc atgggatatc cttagccctc aattcatgta tgggtcaaag
     2641 gcttatgtca agcaccccgc cgacattcca gactatctaa agttatcttt tcccgaaggg
     2701 tttaagtggg agcgtgtgat gaacttcgaa gacggtggcg tggtaacagt gactcaggat
     2761 tcgtccctgc aagatggtga atttatctac aaagtcaaat taagaggaac taactttcca
     2821 tctgacggcc cggttatgca aaaaaagaca atgggctggg aggcctcctc agaacgaatg
     2881 taccctgaag atggtgcctt gaagggtgag attaaacaaa gattgaaatt gaaagatggt
     2941 ggacattatg acgctgaggt taaaacgaca tacaaagcta agaaacctgt ccagctccca
     3001 ggtgcttaca atgtaaatat aaaacttgat attacatcac ataatgaaga ttatacgata
     3061 gttgaacaat acgaaagggc tgaggggaga catagtactg gtggcatgga tgaactatac
     3121 aaataataag cggccgcgag ggccgcatca tgtaattagt tatgtcacgc ttacattcac
     3181 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     3241 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     3301 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     3361 agaaggtttt gggacgctcg aaggctttaa tttgcggcct ctagacgcct tgagcctggc
     3421 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     3481 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     3541 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     3601 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     3661 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     3721 ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc
     3781 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     3841 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     3901 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtcatcga
     3961 tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat
     4021 cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct
     4081 agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt
     4141 ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt
     4201 tttttttgat ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc
     4261 ttcaatttct caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt
     4321 gctcattaga aagaaagcat agcaatctaa tctaagtttt gggatccccc gggctgcagg
     4381 aattcgatat caagcttgga gatctaaaag aaataatgtc taaaggtgaa gaattattca
     4441 ctggtgttgt cccaattttg gttgaattag atggtgatgt taatggtcac aaattttctg
     4501 tctccggtga aggtgaaggt gatgctactt acggtaaatt gaccttaaaa tttatttgta
     4561 ctactggtaa attgccagtt ccatggccaa ccttagtcac tactttcggt tatggtgttc
     4621 aatgttttgc gagataccca gatcatatga aacaacatga ctttttcaag tctgccatgc
     4681 cagaaggtta tgttcaagaa agaactattt ttttcaaaga tgacggtaac tacaagacca
     4741 gagctgaagt caagtttgaa ggtgatacct tagttaatag aatcgaatta aaaggtattg
     4801 attttaaaga agatggtaac attttaggtc acaaattgga atacaactat aactctcaca
     4861 atgtttacat catggctgac aaacaaaaga atggtatcaa agttaacttc aaaattagac
     4921 acaacattga agatggttct gttcaattag ctgaccatta tcaacaaaat actccaattg
     4981 gtgatggtcc agtcttgtta ccagacaacc attacttatc cactcaatct gccttatcca
     5041 aagatccaaa cgaaaagaga gaccacatgg tcttgttaga atttgttact gctgctggta
     5101 ttatccatgg tatggatgaa ttgtacaaat aaagcctagg aaacaaacaa agctgtcacg
     5161 gagtggggtt ccggtctgat gagatcctca tgtgttgacg actcgctgag gatcgaaaca
     5221 gcaaaaagaa aaataaaaac tcgagagggc gcgccacttc taaataagcg aatttcttat
     5281 gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa
     5341 gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca
     5401 ggttgctttc tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat
     5461 ccgctaggga taacagggta atataggtac cagcttttgt tccctttagt gagggttaat
     5521 ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5581 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5641 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5701 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5761 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5821 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5881 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5941 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6061 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6181 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6241 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6301 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6361 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6421 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6481 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6541 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6601 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6661 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6721 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6781 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6841 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6901 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6961 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7021 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7081 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7141 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7201 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7261 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7321 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7381 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7441 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7501 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7561 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7621 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7681 aaaagtgcca cctgggtcct tttcatcacg tgctataaaa ataattataa tttaaatttt
     7741 ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt
     7801 ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat
     7861 cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga
     7921 ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat
     7981 ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt
     8041 ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact
     8101 ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat
     8161 tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatccg tctaagaaac
     8221 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
