LOCUS       sequence-424938-        5795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy plasmid expressing mTurquoise2 under the TDH3p promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5795)
  AUTHORS   Mormino M, Lenitz I, Siewers V, Nygard Y
  TITLE     Identification of acetic acid sensitive strains through 
            biosensor-based screening of a Saccharomyces cerevisiae CRISPRi 
            library.
  JOURNAL   Microb Cell Fact. 2022 Oct 15;21(1):214. doi: 
            10.1186/s12934-022-01938-7.
  PUBMED    36243715
REFERENCE   2  (bases 1 to 5795)
  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: "Microb Cell
            Fact."; date: "2022-10-15"; volume: "21(1):214. doi"; pages: " 
            10.1186/s12934-022-01938-7"
COMMENT     Sequence Label: IL1_1
FEATURES             Location/Qualifiers
     source          1..5795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        212..878
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     748..771
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     terminator      1617..1804
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1672..1696)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            2058..3413
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     CDS             2402..3211
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2469..2488)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3216..3413
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    3418..3921
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(join(4032..4823,4824..4892))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4655..4674
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4893..4997)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5100..5688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5589..5608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagaa cggggtcatc acggctcatc atgcgccaaa caaatgtgtg
       61 caatacacgc tcggatgact gcatgatgac cgcactgact ggggacagca gatccaccta
      121 agcctgtgag agaagcagac acccgacaga tcaaggcagt tacgtctcac caaaccagat
      181 gtcaacacag ctacaacgca gttcgagttt atcattatca atactgccat ttcaaagaat
      241 acgtaaataa ttaatagtag tgattttcct aactttattt agtcaaaaaa ttagcctttt
      301 aattctgctg taacccgtac atgcccaaaa tagggggcgg gttacacaga atatataaca
      361 tcgtaggtgt ctgggtgaac agtttattcc tggcatccac taaatataat ggagcccgct
      421 ttttaagctg gcatccagaa aaaaaaagaa tcccagcacc aaaatattgt tttcttcacc
      481 aaccatcagt tcataggtcc attctcttag cgcaactaca gagaacaggg gcacaaacag
      541 gcaaaaaacg ggcacaacct caatggagtg atgcaacctg cctggagtaa atgatgacac
      601 aaggcaattg acccacgcat gtatctatct cattttctta caccttctat taccttctgc
      661 tctctctgat ttggaaaaag ctgaaaaaaa aggttgaaac cagttccctg aaattattcc
      721 cctacttgac taataagtat ataaagacgg taggtattga ttgtaattct gtaaatctat
      781 ttcttaaact tcttaaattc tacttttata gttagtcttt tttttagttt taaaacacca
      841 agaacttagt ttcgaataaa cacacataaa caaacaaaag atctatggtt tctaaaggtg
      901 aagaattatt cactggtgtt gtcccaattt tggttgaatt agatggtgat gttaatggtc
      961 acaaattttc tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa ttgaccttaa
     1021 aatttatttg tactactggt aaattgccag ttccatggcc aaccttagtc actactttat
     1081 cttggggtgt tcaatgtttt gcaagatacc cagatcatat gaaacaacat gactttttca
     1141 agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa gatgacggta
     1201 actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat agaatcgaat
     1261 taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg gaatacaatt
     1321 atttctctga caatgtttac atcactgctg acaaacaaaa gaatggtatc aaagctaact
     1381 tcaaaattag acacaacatt gaagatggtg gtgttcaatt agctgaccat tatcaacaaa
     1441 atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta tccactcaat
     1501 ctaagttatc caaagatcca aacgaaaaga gggaccacat ggtcttgtta gaatttgtta
     1561 ctgctgctgg tattaccttg ggtatggatg aattgtacaa aggatcctaa ctcgaggcga
     1621 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
     1681 aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc
     1741 tgtaggtcag gttgctttct caggtatagc atgaggtcgc tcttattgac cacacctcta
     1801 ccggcatgcc gagcaaatgc ctgcaaatcg ctccccattt cgctgccaat gagacgacgg
     1861 ggtcatcacg gctcatcatg cgccaaacaa atgtgtgcaa tacacgctcg gatgactgca
     1921 tgatgaccgc actgactggg gacagcagat ccacctaagc ctgtgagaga agcagacacc
     1981 cgacagatca aggcagttaa ctagtgcact gcagtacagt ttagcttgcc tcgtccccgc
     2041 cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt cttgacgtgc
     2101 gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat
     2161 aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct
     2221 cgctccagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg
     2281 ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata
     2341 tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaaa
     2401 aatgggtaag gaaaagactc acgtttcgag gccgcgatta aattccaaca tggatgctga
     2461 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2521 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2581 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2641 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2701 cggcaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2761 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2821 cagcgatcgc gtatttcgtc tggctcaggc gcaatcacga atgaataacg gtttggttga
     2881 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2941 gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3001 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3061 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3121 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3181 gtttcatttg atgctcgatg agtttttcta aagtaactga caataaaaag attcttgttt
     3241 tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg
     3301 ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg
     3361 cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctggagtatc
     3421 acgtgctata aaaataatta taatttaaat tttttaatat aaatatataa attaaaaata
     3481 gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt ttccgaagat gtaaaagact
     3541 ctagggggat cgccaacaaa tactaccttt tatcttgctc ttcctgctct caggtattaa
     3601 tgccgaattg tttcatcttg tctgtgtaga aaaccacaca cgaaaatcct gtgattttac
     3661 attttactta tcgttaatcg aatgtatatc tatttaatct gcttttcttg tctaataaat
     3721 atatatgtaa agtacgcttt ttgttgaaat tttttaaacc tttgtttatt tttttttctt
     3781 cattccgtaa ctcttctacc ttctttattt actttctaaa atccaaatac aaaacataaa
     3841 aataaataaa cacagagtaa attcccaaat tattccatca ttaaaagata cgaggcgcgt
     3901 gtaagttaca ggcaagcgat ccgtcccgag cggccgcgat tatcaaaaag gatcttcacc
     3961 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4021 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4081 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4141 ccatctggcc ccagtgctgc aatgataccg cgggacccac gctcaccggc tccagattta
     4201 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4261 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4321 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4381 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4441 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4501 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4561 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4621 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4681 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4741 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4801 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4861 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4921 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4981 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgtcatgac caaaatccct
     5041 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5101 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5161 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5221 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5281 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5341 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5401 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5461 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5521 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5581 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5641 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5701 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5761 ttatcccctg attctgtgga taaccgtgcg gccgc
//
