LOCUS       sequence-351961-        5366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genome integration in X2 site expressing OxPro 
            (oxidative stress biosensor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5366)
  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 5366)
  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_7_OxPro
FEATURES             Location/Qualifiers
     source          1..5366
                     /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"
     primer_bind     complement(1670..1686)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1704..1722)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     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"
     CDS             2367..3077
                     /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(2512..2530)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2780..2799
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3091..3278
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3146..3170)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3527..4342)
                     /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(3527..4342)
                     /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     4250..4269
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4334..4353
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4567..5155
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5056..5075
                     /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 tgagcaaggg cgaggaggat aacatggcca
     2401 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtggac ggccacgagt
     2461 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     2521 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     2581 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     2641 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     2701 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     2761 ccaacttccc ctccgacggc cccgtaatgc agaagaagac tatgggctgg gaggcctcct
     2821 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     2881 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     2941 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     3001 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     3061 acgagctgta caagtaaatc ctaactcgag gcgaatttct tatgatttat gatttttatt
     3121 attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta
     3181 aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta
     3241 tagcatgagg tcgctcttat tgaccacacc tctaccggca tgccgagcaa atgcctgcaa
     3301 atcgctcccc atttcgctga gcagttacag agatgttacg aaccactagt gcactgcagt
     3361 acataacccc taggtcttcc tgagagtgac gaaggctaag gtcacttctt cgtttccttt
     3421 attggggttt ccgtgtagcc ttcccctgaa tagtgtggga cgttttatga gaagccgtaa
     3481 gaaatagccg agcggccgcg tgttacaacc aattaaccaa ttctgattag aaaaactcat
     3541 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     3601 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     3661 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     3721 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     3781 atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     3841 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgaggc
     3901 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     3961 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     4021 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     4081 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     4141 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     4201 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     4261 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctg gagcaagacg
     4321 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     4381 ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     4441 caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag tcatgaccaa
     4501 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4561 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4621 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4681 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4741 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4801 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4861 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4921 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4981 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5041 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5101 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5161 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5221 tcctgcgtta tcccctgatt ctgtggataa ccgtgcggcc gccaatgtca aaagatcctc
     5281 tcataccata ttaagtaaat tgcctccatt tctttttcct cgggcagaga aactcgcagg
     5341 caacttgctc tcgaagtggt cacgtg
//
