LOCUS       sequence-378782-        6932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genome integration in X2 site expressing GlyRNA and 
            OxPro (biosensors for glycolytic flux and oxidative stress).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6932)
  AUTHORS   Torello Pianale L, Rugbjerg P, Olsson L
  TITLE     Real-Time Monitoring of the Yeast Intracellular State During 
            Bioprocesses With a Toolbox of Biosensors.
  JOURNAL   Front Microbiol. 2022 Jan 7;12:802169. doi: 
            10.3389/fmicb.2021.802169. eCollection 2021.
  PUBMED    35069506
REFERENCE   2  (bases 1 to 6932)
  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: "Front 
            Microbiol."; date: "2022-01-7"; volume: "12:802169. doi"; pages: " 
            10.3389/fmicb.2021.802169. eCollection 2021"
COMMENT     Sequence Label: LT2_14_GlyOx
FEATURES             Location/Qualifiers
     source          1..6932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1669..1685)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1956..2353
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             complement(2252..2263)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3235..3251)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3522..3919
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             complement(3818..3829)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3933..4643
                     /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(4078..4096)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4346..4365
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      4657..4844
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4712..4736)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(5093..5908)
                     /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(5093..5908)
                     /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     5816..5835
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5900..5919
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6133..6721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6622..6641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagac tgaactggcc gataattgca gacgaacgaa cggaatgcgt
       61 gcgatcttga gtgccagtga atattaagta atcatcaaag tatatgtgta attgtttata
      121 ctcttagtaa aggatgctcc ctacaaggtg gctcttttct tactaagcgc gttcagtttc
      181 cagccagccg aaagagggat atcagcttga gtgccagtga atattaagta atcatcaaag
      241 tatatgtgta attgtttata ctcttagtaa aggatgctcc ctacaaggtg gctcttttct
      301 tactaagcgc gttcagtttc cagccagccg aaagagggat atcagacctg cactcttgag
      361 tgccagtgaa tattaagtaa tcatcaaagt atatgtgtaa ttgtttatac tcttagtaaa
      421 ggatgctccc tacaaggtgg ctcttttctt actaagcgcg ttcagtttcc agccagccga
      481 aagagggata tcagcttgag tgccagtgaa tattaagtaa tcatcaaagt atatgtgtaa
      541 ttgtttatac tcttagtaaa ggatgctccc tacaaggtgg ctcttttctt actaagcgcg
      601 ttcagtttcc agccagccga aagagggata tcagatctgt catcttgagt gccagtgaat
      661 attaagtaat catcaaagta tatgtgtaat tgtttatact cttagtaaag gatgctccct
      721 acaaggtggc tcttttctta ctaagcgcgt tcagtttcca gccagccgaa agagggatat
      781 cagtatataa gaaagccatt cgggggatga aaagctgaca agagaataac gaggaccagt
      841 ttttatttgt tgtctagcaa gaattataca cgcacacata cacgagagtc tacgatatct
      901 ttaaataaca catcaataat ctgtcataaa acatatggta tccaagggcg aagaattgtt
      961 caccggtgtg gtcccaatct tggtcgagtt ggacggtgac gtcaacggtc acaagttctc
     1021 tgtttccggt gaaggtgaag gagatgccac ctacggtaaa ttgaccttga agttgttgtg
     1081 caccaccggc aagctgccag ttccatggcc aacattggtc accactttag gttatggttt
     1141 acaatgtttt gctagatatc cagatcatat gaaacaacat gatttcttca aatctgctat
     1201 gccagaaggt tatgttcaag aaagaactat atttttcaaa gatgatggta attataaaac
     1261 tagagctgaa gttaaatttg aaggagatac tttagttaat agaattgaat tgaaaggtat
     1321 tgattttaaa gaagatggta atattttagg tcataaatta gaatataatt ataattctca
     1381 taatgtttat attacagctg ataaacaaaa gaatggtatt aaggctaatt ttaaaattag
     1441 acataatatt gaagatggtg gtgttcaatt agctgatcat tatcaacaaa atactcctat
     1501 tggtgacggt ccagttttat tacctgataa tcattatttg tcatatcaat ctaaattgtc
     1561 aaaagatcca aatgaaaaaa gagatcatat ggttttgtta gaatttttga ctgcagctgg
     1621 tattacttta ggtatggatg aattgtataa aggatcctaa ctcgagatcg ataccgtcga
     1681 cctcgagtca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct
     1741 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa
     1801 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     1861 aacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct
     1921 cggctgccaa accagatgtc aacacagcta caacgatagc ttcgaaacgt ttctactcct
     1981 cttttactct tccagatctt ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca
     2041 agcacagcat actaaatttc ccctctttct tcctctaggg tgtcgttaat tacccgtact
     2101 aaaggtttgg aaaagaaaaa agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa
     2161 taaaaatttt tatcacgttt ctttttcttg aaaattcttt ttttgatttt tttctctttc
     2221 gatgacctcc catcgatatt taagttaata aacggccttc aatttcccaa gtttcagtcc
     2281 cacttttctt gctctactac aacttttttt acctcttgct cattagaaag aaagcatagc
     2341 aatctaatct aagttttcta gaaaatatgg tttctaaagg tgaagaatta ttcactggtg
     2401 ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt tctgtctccg
     2461 gtgaaggtga aggtgatgct acttacggta aattgacctt aaaatttatt tgtactactg
     2521 gtaaattgcc agttccatgg ccaaccttag tcactacttt atcttggggt gttcaatgtt
     2581 ttgcaagata cccagatcat atgaaacaac atgacttttt caagtctgcc atgccagaag
     2641 gttatgttca agaaagaact atttttttca aagatgacgg taactacaag accagagctg
     2701 aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt attgatttta
     2761 aagaagatgg taacatttta ggtcacaaat tggaatacaa ttatttctct gacaatgttt
     2821 acatcactgc tgacaaacaa aagaatggta tcaaagctaa cttcaaaatt agacacaaca
     2881 ttgaagatgg tggtgttcaa ttagctgacc attatcaaca aaatactcca attggtgatg
     2941 gtccagtctt gttaccagac aaccattact tatccactca atctaagtta tccaaagatc
     3001 caaacgaaaa gagggaccac atggtcttgt tagaatttgt tactgctgct ggtattacct
     3061 tgggtatgga tgaattgtac aaaggatcct aaagcctagg aaacaaacaa agctgtcacc
     3121 ggaataggag tccggtctga tgagtccact tgctgaggat ccgacggttc ctcatgtgtt
     3181 gacgaaggac gaaacagcaa aaagaaaaat aaaaactcga gcctcgagtg gcatcgatac
     3241 cgtcgacctc gagtcatgta attagttatg tcacgcttac attcacgccc tcctcccaca
     3301 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     3361 ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     3421 tgtacaaacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg
     3481 gacgctcggc tggatgcaca cactggctta agatgacaac gatagcttcg aaacgtttct
     3541 actcctcttt tactcttcca gatcttctcg gactccgcgc atcgccgtac cacttcaaaa
     3601 cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc gttaattacc
     3661 cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa
     3721 aggcaataaa aatttttatc acgtttcttt ttcttgaaaa ttcttttttt gatttttttc
     3781 tctttcgatg acctcccatc gatatttaag ttaataaacg gccttcaatt tcccaagttt
     3841 cagtcccact tttcttgctc tactacaact ttttttacct cttgctcatt agaaagaaag
     3901 catagcaatc taatctaagt tttctagaaa atatggtgag caagggcgag gaggataaca
     3961 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtggacggcc
     4021 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     4081 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     4141 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     4201 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     4261 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     4321 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagactatg ggctgggagg
     4381 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     4441 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     4501 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     4561 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     4621 gcatggacga gctgtacaag taaatcctaa ctcgaggcga atttcttatg atttatgatt
     4681 tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag
     4741 gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct
     4801 caggtatagc atgaggtcgc tcttattgac cacacctcta ccggcatgcc gagcaaatgc
     4861 ctgcaaatcg ctccccattt cgctgagcag ttacagagat gttacgaacc actagtgcac
     4921 tgcagtacat aacccctagg tcttcctgag agtgacgaag gctaaggtca cttcttcgtt
     4981 tcctttattg gggtttccgt gtagccttcc cctgaatagt gtgggacgtt ttatgagaag
     5041 ccgtaagaaa tagccgagcg gccgcgtgtt acaaccaatt aaccaattct gattagaaaa
     5101 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     5161 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     5221 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     5281 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     5341 gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac
     5401 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     5461 cgaggcgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     5521 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     5581 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     5641 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     5701 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     5761 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     5821 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctggagc
     5881 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     5941 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     6001 gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagtcat
     6061 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6121 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6181 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6241 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6301 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6361 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6421 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6481 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6541 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6601 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6661 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6721 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6781 gttctttcct gcgttatccc ctgattctgt ggataaccgt gcggccgcca atgtcaaaag
     6841 atcctctcat accatattaa gtaaattgcc tccatttctt tttcctcggg cagagaaact
     6901 cgcaggcaac ttgctctcga agtggtcacg tg
//
