LOCUS       sequence-384687-        5092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genome integration in X2 site expressing PyruPro 
            (Pyruvate consumption Probe).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5092)
  AUTHORS   Torello Pianale L, Olsson L
  TITLE     ScEnSor Kit for Saccharomyces cerevisiae Engineering and 
            Biosensor-Driven Investigation of the Intracellular Environment.
  JOURNAL   ACS Synth Biol. 2023 Aug 8. doi: 10.1021/acssynbio.3c00124.
  PUBMED    37552581
REFERENCE   2  (bases 1 to 5092)
  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: "ACS Synth 
            Biol. 2023 Aug 8. doi: 10.1021/acssynbio.3c00124."
COMMENT     Sequence Label: LT2_33_PyruPro_X2site
FEATURES             Location/Qualifiers
     source          1..5092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1389..1405)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1390..1406)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1424..1442)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        1676..2073
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             complement(1972..1983)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2087..2797
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVDGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2232..2250)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2500..2519
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2811..2998
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2866..2890)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3247..4062)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3247..4062)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3970..3989
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4054..4073
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4287..4875
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4776..4795
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attcgcatct agactgaact ggccgataat tgcagacgaa cggtagcttc cagtttttgg
       61 tggttctggc ttccttcccg attccgcccg ctaaacgcat aattttgttg cctggtggca
      121 tttgcaaaat gcataaccta tgcatttaaa agattatgta tgctcttctg acttttcgtg
      181 tgatgaggct cgtggaaaaa atgaataatt tatgaatttg agaacaattt tgtgttgtta
      241 cggtatttta ctatggaata atcaatcaat tgaggatttt atgcaaatat cgtttgaata
      301 tttttccgac cctttgagta cttttcttca taattgcata atattgtccg ctgcccgttt
      361 ttctgttaga cggtgtcttg atctacttgc tatcgttcaa caccacctta ttttctaact
      421 attttttttt tagctcattt gaatcagctt atggtgatgg cacatttttg cataaaccta
      481 gctgtcctcg ttgaacatag gaaaaaaaaa tatataaaca aggctctttc actctccttg
      541 gaatcagatt tgggtttgtt ccctttattt tcatatttct tgtcatattc ttttctcaat
      601 tattatcttc tactcataac ctcacgcaaa ataacacagt caaatcaatc aaatatggta
      661 tccaagggcg aagaattgtt caccggtgtg gtcccaatct tggtcgagtt ggacggtgac
      721 gtcaacggtc acaagttctc tgtttccggt gaaggtgaag gagatgccac ctacggtaaa
      781 ttgaccttga agttgttgtg caccaccggc aagctgccag ttccatggcc aacattggtc
      841 accactttag gttatggttt acaatgtttt gctagatatc cagatcatat gaaacaacat
      901 gatttcttca aatctgctat gccagaaggt tatgttcaag aaagaactat atttttcaaa
      961 gatgatggta attataaaac tagagctgaa gttaaatttg aaggagatac tttagttaat
     1021 agaattgaat tgaaaggtat tgattttaaa gaagatggta atattttagg tcataaatta
     1081 gaatataatt ataattctca taatgtttat attacagctg ataaacaaaa gaatggtatt
     1141 aaggctaatt ttaaaattag acataatatt gaagatggtg gtgttcaatt agctgatcat
     1201 tatcaacaaa atactcctat tggtgacggt ccagttttat tacctgataa tcattatttg
     1261 tcatatcaat ctaaattgtc aaaagatcca aatgaaaaaa gagatcatat ggttttgtta
     1321 gaatttttga ctgcagctgg tattacttta ggtatggatg aattgtataa aggatcctaa
     1381 ctcgagatcg ataccgtcga cctcgagtca tgtaattagt tatgtcacgc ttacattcac
     1441 gccctcctcc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     1501 tccctattta ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat
     1561 ttttcttttt tttctgtaca aacgcgtgta cgcatgtaac attatactga aaaccttgct
     1621 tgagaaggtt ttgggacgct cggctgccaa accagatgtc aacacagcta caacgatagc
     1681 ttcgaaacgt ttctactcct cttttactct tccagatctt ctcggactcc gcgcatcgcc
     1741 gtaccacttc aaaacaccca agcacagcat actaaatttc ccctctttct tcctctaggg
     1801 tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa agagaccgcc tcgtttcttt
     1861 ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt ctttttcttg aaaattcttt
     1921 ttttgatttt tttctctttc gatgacctcc catcgatatt taagttaata aacggccttc
     1981 aatttcccaa gtttcagtcc cacttttctt gctctactac aacttttttt acctcttgct
     2041 cattagaaag aaagcatagc aatctaatct aagttttcta gaaaatatgg tgagcaaggg
     2101 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     2161 ctccgtggac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     2221 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     2281 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     2341 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     2401 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     2461 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     2521 tatgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     2581 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     2641 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     2701 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     2761 ccactccacc ggcggcatgg acgagctgta caagtaaatc ctaactcgag gcgaatttct
     2821 tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt
     2881 aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg
     2941 tcaggttgct ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggca
     3001 tgccgagcaa atgcctgcaa atcgctcccc atttcgctga gcagttacag agatgttacg
     3061 aaccactagt gcactgcagt acataacccc taggtcttcc tgagagtgac gaaggctaag
     3121 gtcacttctt cgtttccttt attggggttt ccgtgtagcc ttcccctgaa tagtgtggga
     3181 cgttttatga gaagccgtaa gaaatagccg agcggccgcg tgttacaacc aattaaccaa
     3241 ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     3301 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     3361 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     3421 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     3481 gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac
     3541 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     3601 tgattgcgcc tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     3661 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     3721 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     3781 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     3841 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     3901 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     3961 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     4021 tcgcggcctg gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     4081 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     4141 acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt
     4201 gaaggatcag tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4261 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4321 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4381 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4441 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4501 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4561 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4621 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4681 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4741 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4801 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4861 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4921 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtgcggcc
     4981 gccaatgtca aaagatcctc tcataccata ttaagtaaat tgcctccatt tctttttcct
     5041 cgggcagaga aactcgcagg caacttgctc tcgaagtggt cacgtgccct ga
//
