LOCUS       sequence-351964-        5025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genome integration in X2 site expressing GlyRNA 
            (glycolytic flux biosensor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5025)
  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 5025)
  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_12_GlyRNA
FEATURES             Location/Qualifiers
     source          1..5025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        49..446
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             complement(345..356)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1328..1344)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1329..1345)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1363..1381)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        1615..2012
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             complement(1911..1922)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2026..2736
                     /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(2171..2189)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2439..2458
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2750..2937
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2805..2829)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3186..4001)
                     /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(3186..4001)
                     /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     3909..3928
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3993..4012
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4226..4814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4715..4734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagac tgaactggcc gataattgca gacgaacgat agcttcgaaa
       61 cgtttctact cctcttttac tcttccagat cttctcggac tccgcgcatc gccgtaccac
      121 ttcaaaacac ccaagcacag catactaaat ttcccctctt tcttcctcta gggtgtcgtt
      181 aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg
      241 tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaaattc tttttttgat
      301 ttttttctct ttcgatgacc tcccatcgat atttaagtta ataaacggcc ttcaatttcc
      361 caagtttcag tcccactttt cttgctctac tacaactttt tttacctctt gctcattaga
      421 aagaaagcat agcaatctaa tctaagtttt ctagaaaata tggtttctaa aggtgaagaa
      481 ttattcactg gtgttgtccc aattttggtt gaattagatg gtgatgttaa tggtcacaaa
      541 ttttctgtct ccggtgaagg tgaaggtgat gctacttacg gtaaattgac cttaaaattt
      601 atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttatcttgg
      661 ggtgttcaat gttttgcaag atacccagat catatgaaac aacatgactt tttcaagtct
      721 gccatgccag aaggttatgt tcaagaaaga actatttttt tcaaagatga cggtaactac
      781 aagaccagag ctgaagtcaa gtttgaaggt gataccttag ttaatagaat cgaattaaaa
      841 ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca aattggaata caattatttc
      901 tctgacaatg tttacatcac tgctgacaaa caaaagaatg gtatcaaagc taacttcaaa
      961 attagacaca acattgaaga tggtggtgtt caattagctg accattatca acaaaatact
     1021 ccaattggtg atggtccagt cttgttacca gacaaccatt acttatccac tcaatctaag
     1081 ttatccaaag atccaaacga aaagagggac cacatggtct tgttagaatt tgttactgct
     1141 gctggtatta ccttgggtat ggatgaattg tacaaaggat cctaaagcct aggaaacaaa
     1201 caaagctgtc accggaatag gagtccggtc tgatgagtcc acttgctgag gatccgacgg
     1261 ttcctcatgt gttgacgaag gacgaaacag caaaaagaaa aataaaaact cgagcctcga
     1321 gtggcatcga taccgtcgac ctcgagtcat gtaattagtt atgtcacgct tacattcacg
     1381 ccctcctccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
     1441 ccctatttat tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt
     1501 tttctttttt ttctgtacaa acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
     1561 gagaaggttt tgggacgctc ggctgccaaa ccagatgtca acacagctac aacgatagct
     1621 tcgaaacgtt tctactcctc ttttactctt ccagatcttc tcggactccg cgcatcgccg
     1681 taccacttca aaacacccaa gcacagcata ctaaatttcc cctctttctt cctctagggt
     1741 gtcgttaatt acccgtacta aaggtttgga aaagaaaaaa gagaccgcct cgtttctttt
     1801 tcttcgtcga aaaaggcaat aaaaattttt atcacgtttc tttttcttga aaattctttt
     1861 tttgattttt ttctctttcg atgacctccc atcgatattt aagttaataa acggccttca
     1921 atttcccaag tttcagtccc acttttcttg ctctactaca acttttttta cctcttgctc
     1981 attagaaaga aagcatagca atctaatcta agttttctag aaaatatggt gagcaagggc
     2041 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     2101 tccgtggacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     2161 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     2221 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     2281 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     2341 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     2401 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagact
     2461 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     2521 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     2581 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     2641 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     2701 cactccaccg gcggcatgga cgagctgtac aagtaaatcc taactcgagg cgaatttctt
     2761 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     2821 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     2881 caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat
     2941 gccgagcaaa tgcctgcaaa tcgctcccca tttcgctgag cagttacaga gatgttacga
     3001 accactagtg cactgcagta cataacccct aggtcttcct gagagtgacg aaggctaagg
     3061 tcacttcttc gtttccttta ttggggtttc cgtgtagcct tcccctgaat agtgtgggac
     3121 gttttatgag aagccgtaag aaatagccga gcggccgcgt gttacaacca attaaccaat
     3181 tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     3241 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     3301 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     3361 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     3421 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca
     3481 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     3541 gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     3601 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     3661 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     3721 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     3781 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     3841 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     3901 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     3961 cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     4021 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     4081 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
     4141 aaggatcagt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4201 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4261 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4321 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4381 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4441 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4501 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4561 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4621 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4681 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4741 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4801 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4861 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtgcggccg
     4921 ccaatgtcaa aagatcctct cataccatat taagtaaatt gcctccattt ctttttcctc
     4981 gggcagagaa actcgcaggc aacttgctct cgaagtggtc acgtg
//
