LOCUS       sequence-384739-        4579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genome integration in X2 site expressing UPRpro - Sensor
            for Unfolded Protein Response.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4579)
  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 4579)
  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_15_UPRpro_X2site
FEATURES             Location/Qualifiers
     source          1..4579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(883..899)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(884..900)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(918..936)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        1170..1567
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             complement(1466..1477)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1581..2291
                     /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(1726..1744)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1994..2013
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2305..2492
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2360..2384)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2741..3556)
                     /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(2741..3556)
                     /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     3464..3483
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3548..3567
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3781..4369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4270..4289
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccctgaatt cgcatctaga ctgaactggc cgataattgc agacgaacgg gacagcgtgt
       61 ccttaagatc ttgtaatatt ctaatcaagc ttataaaaga gcactgttgg gcgtgagtgg
      121 aggcgccgga ataaaccatg caatgaatat ggtatccaag ggcgaagaat tgttcaccgg
      181 tgtggtccca atcttggtcg agttggacgg tgacgtcaac ggtcacaagt tctctgtttc
      241 cggtgaaggt gaaggagatg ccacctacgg taaattgacc ttgaagttgt tgtgcaccac
      301 cggcaagctg ccagttccat ggccaacatt ggtcaccact ttaggttatg gtttacaatg
      361 ttttgctaga tatccagatc atatgaaaca acatgatttc ttcaaatctg ctatgccaga
      421 aggttatgtt caagaaagaa ctatattttt caaagatgat ggtaattata aaactagagc
      481 tgaagttaaa tttgaaggag atactttagt taatagaatt gaattgaaag gtattgattt
      541 taaagaagat ggtaatattt taggtcataa attagaatat aattataatt ctcataatgt
      601 ttatattaca gctgataaac aaaagaatgg tattaaggct aattttaaaa ttagacataa
      661 tattgaagat ggtggtgttc aattagctga tcattatcaa caaaatactc ctattggtga
      721 cggtccagtt ttattacctg ataatcatta tttgtcatat caatctaaat tgtcaaaaga
      781 tccaaatgaa aaaagagatc atatggtttt gttagaattt ttgactgcag ctggtattac
      841 tttaggtatg gatgaattgt ataaaggatc ctaactcgag atcgataccg tcgacctcga
      901 gtcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc
      961 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta
     1021 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaacgcg
     1081 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcggctg
     1141 ccaaaccaga tgtcaacaca gctacaacga tagcttcgaa acgtttctac tcctctttta
     1201 ctcttccaga tcttctcgga ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca
     1261 gcatactaaa tttcccctct ttcttcctct agggtgtcgt taattacccg tactaaaggt
     1321 ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa
     1381 tttttatcac gtttcttttt cttgaaaatt ctttttttga tttttttctc tttcgatgac
     1441 ctcccatcga tatttaagtt aataaacggc cttcaatttc ccaagtttca gtcccacttt
     1501 tcttgctcta ctacaacttt ttttacctct tgctcattag aaagaaagca tagcaatcta
     1561 atctaagttt tctagaaaat atggtgagca agggcgagga ggataacatg gccatcatca
     1621 aggagttcat gcgcttcaag gtgcacatgg agggctccgt ggacggccac gagttcgaga
     1681 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     1741 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     1801 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     1861 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     1921 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     1981 tcccctccga cggccccgta atgcagaaga agactatggg ctgggaggcc tcctccgagc
     2041 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     2101 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     2161 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     2221 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     2281 tgtacaagta aatcctaact cgaggcgaat ttcttatgat ttatgatttt tattattaaa
     2341 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
     2401 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat
     2461 gaggtcgctc ttattgacca cacctctacc ggcatgccga gcaaatgcct gcaaatcgct
     2521 ccccatttcg ctgagcagtt acagagatgt tacgaaccac tagtgcactg cagtacataa
     2581 cccctaggtc ttcctgagag tgacgaaggc taaggtcact tcttcgtttc ctttattggg
     2641 gtttccgtgt agccttcccc tgaatagtgt gggacgtttt atgagaagcc gtaagaaata
     2701 gccgagcggc cgcgtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca
     2761 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     2821 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     2881 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     2941 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     3001 aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     3061 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg aggcgaaata
     3121 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     3181 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     3241 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     3301 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     3361 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     3421 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     3481 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctggagcaa gacgtttccc
     3541 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     3601 ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt
     3661 ggctttgttg aataaatcga acttttgctg agttgaagga tcagtcatga ccaaaatccc
     3721 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3781 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3841 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3901 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3961 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4021 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4081 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4141 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4201 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4261 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4321 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4381 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4441 gttatcccct gattctgtgg ataaccgtgc ggccgccaat gtcaaaagat cctctcatac
     4501 catattaagt aaattgcctc catttctttt tcctcgggca gagaaactcg caggcaactt
     4561 gctctcgaag tggtcacgt
//
