LOCUS       sequence-384738-        4921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genome integration in X2 site expressing ResPro (Ethanol
            consumption Probe).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4921)
  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 4921)
  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_34_EthPro_X2site
FEATURES             Location/Qualifiers
     source          1..4921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1233..1249)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1234..1250)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1268..1286)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        1520..1917
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             complement(1816..1827)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1931..2641
                     /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(2076..2094)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2344..2363
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2655..2842
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2710..2734)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3091..3906)
                     /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(3091..3906)
                     /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     3814..3833
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3898..3917
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4131..4719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4620..4639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtcacgtgc cctgaattcg catctagact gaactggccg ataattgcag acgaacgtcc
       61 tgcctttcta atcaccattc taatgtttta attaagggat tttgtcttca ttaacggctt
      121 tcgctcataa aaatgttatg acgttttgcc cgcaggcggg aaaccatcca cttcacgaga
      181 ctgatctcct ctgccggaac accgggcatc tccaacttat aagttggaga aataagagaa
      241 tttcagattg agagaatgaa aaaaaaaaaa aaaaaggcag aggagagcat aaaaatgggg
      301 ttcacttttt ggtaaagcta tagcatgcct atcacatata aatagagtgc cagtagcgac
      361 ttttttcaca ctcgaaatac tcttactact gctctcttgt tgtttttatc acttcttgtt
      421 tcttcttggt aaatagaata tcaagctaca aaaagcatac aatcaactat caactattaa
      481 ctatatcgta atacacatat ggtttctaaa ggtgaagaat tattcactgg tgttgtccca
      541 attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt
      601 gaaggtgatg ctacttacgg taaattgacc ttaaaattta tttgtactac tggtaaattg
      661 ccagttccat ggccaacctt agtcactact ttatcttggg gtgttcaatg ttttgcaaga
      721 tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga aggttatgtt
      781 caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc tgaagtcaag
      841 tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt taaagaagat
      901 ggtaacattt taggtcacaa attggaatac aattatttct ctgacaatgt ttacatcact
      961 gctgacaaac aaaagaatgg tatcaaagct aacttcaaaa ttagacacaa cattgaagat
     1021 ggtggtgttc aattagctga ccattatcaa caaaatactc caattggtga tggtccagtc
     1081 ttgttaccag acaaccatta cttatccact caatctaagt tatccaaaga tccaaacgaa
     1141 aagagggacc acatggtctt gttagaattt gttactgctg ctggtattac cttgggtatg
     1201 gatgaattgt acaaaggatc ctaactcgag atcgataccg tcgacctcga gtcatgtaat
     1261 tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag
     1321 gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt
     1381 aagaacgtta tttatatttc aaatttttct tttttttctg tacaaacgcg tgtacgcatg
     1441 taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcggctg ccaaaccaga
     1501 tgtcaacaca gctacaacga tagcttcgaa acgtttctac tcctctttta ctcttccaga
     1561 tcttctcgga ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa
     1621 tttcccctct ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga
     1681 aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac
     1741 gtttcttttt cttgaaaatt ctttttttga tttttttctc tttcgatgac ctcccatcga
     1801 tatttaagtt aataaacggc cttcaatttc ccaagtttca gtcccacttt tcttgctcta
     1861 ctacaacttt ttttacctct tgctcattag aaagaaagca tagcaatcta atctaagttt
     1921 tctagaaaat atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
     1981 gcgcttcaag gtgcacatgg agggctccgt ggacggccac gagttcgaga tcgagggcga
     2041 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     2101 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     2161 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     2221 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     2281 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     2341 cggccccgta atgcagaaga agactatggg ctgggaggcc tcctccgagc ggatgtaccc
     2401 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     2461 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     2521 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     2581 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta
     2641 aatcctaact cgaggcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     2701 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
     2761 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat gaggtcgctc
     2821 ttattgacca cacctctacc ggcatgccga gcaaatgcct gcaaatcgct ccccatttcg
     2881 ctgagcagtt acagagatgt tacgaaccac tagtgcactg cagtacataa cccctaggtc
     2941 ttcctgagag tgacgaaggc taaggtcact tcttcgtttc ctttattggg gtttccgtgt
     3001 agccttcccc tgaatagtgt gggacgtttt atgagaagcc gtaagaaata gccgagcggc
     3061 cgcgtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
     3121 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     3181 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     3241 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     3301 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg
     3361 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     3421 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg aggcgaaata cgcgatcgct
     3481 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     3541 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     3601 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     3661 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     3721 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     3781 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     3841 atcagcatcc atgttggaat ttaatcgcgg cctggagcaa gacgtttccc gttgaatatg
     3901 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     3961 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg
     4021 aataaatcga acttttgctg agttgaagga tcagtcatga ccaaaatccc ttaacgtgag
     4081 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4141 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4201 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4261 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4321 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4381 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4441 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4501 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4561 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4621 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4681 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4741 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4801 gattctgtgg ataaccgtgc ggccgccaat gtcaaaagat cctctcatac catattaagt
     4861 aaattgcctc catttctttt tcctcgggca gagaaactcg caggcaactt gctctcgaag
     4921 t
//
