LOCUS       sequence-497172-        6349 bp    DNA     circular SYN 13-JAN-2026
DEFINITION  mutated LuxR transcriptional activator for expression in yeast: 
            fused with Gal4 activation domain with 'gen2' mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6349)
  AUTHORS   van Aalst ACA, Holtz M, Lyskjaer Jensen M, Schroder T, Crocoll C, 
            Poborsky M, Damgaard Jensen E, Krogh Jensen M
  TITLE     Engineering N-acyl-homoserine lactone-based quorum-sensing circuit 
            for dynamic regulatory control in Saccharomyces cerevisiae.
  JOURNAL   Commun Biol. 2025 Nov 27. doi: 10.1038/s42003-025-09163-9.
  PUBMED    41310402
REFERENCE   2  (bases 1 to 6349)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 13, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol. 2025 Nov 27. doi: 10.1038/s42003-025-09163-9."
COMMENT     Sequence Label: pAvA143
FEATURES             Location/Qualifiers
     source          1..6349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     622..641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(860..951)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     993..1012
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1018..1037
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1244..1277
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             2904..3245
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTEMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     CDS             3255..3275
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      4064..4229
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4119..4143)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4255..4274)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4256..4274)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4284..4300)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4293..4315)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(4849..4882)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(5352..5368)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5467..5484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5638..6226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5718..5737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6348..6349,1..790,791..859))
                     /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"
ORIGIN
        1 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
       61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      121 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
      181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
      241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      301 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      841 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
      901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
      961 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     1021 gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1081 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc ccatgcgaga
     1141 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1201 cccgggctaa ttatggggtg tcgcccttat tcgactctat agtgaagttc ctattctcta
     1261 gaaagtatag gaacttctga agtggggatt taaatgcggc cgcgctgagg gtttaatccc
     1321 aaagctaaga gtcccatttt attcttctat atgtatattt tcgatactct aaaccaccct
     1381 acaatgtagc cctatactaa atctgctcaa ttttcagctt ctacaagtga ctcgaggcca
     1441 cgtggaaaga tccaactact ccagcacaac gattcaatat aatcgattgc tccactcata
     1501 agaggcaaga acaagcttca acttttggta agccgccgtt tataaacagg gaagatgtcc
     1561 tttgtcaagg gaggcacaga gcatggccaa tttggcaaat tgcaggtttt tctgagtgaa
     1621 aaatgaaaaa gcattgtagt agagtcggct cactgaaaaa ccggggagga cgaaaaggtt
     1681 tccagccaca gttgtagtca cgtgcgcgcc atgctgacta atggcagccg tcgttgggca
     1741 gaagagaatt agtatggtac aggatacgct aattgcgctc caactaccaa ggttgttgag
     1801 ggaacactgg ggcaataggc tgtcgccatt caagagcaga ttaagtcctc agcgagctcg
     1861 catggaatgc gtgcgatagt gcgatagacg cgaatttttc gaagaagtac cttcaaagaa
     1921 tggggtctta tcttgttttg caagtaccac tgagcaggat aataatagaa atgataatat
     1981 actatagtag agataacgtc gatgacttcc catactgtaa ttgcttttag ttgtgtattt
     2041 ttagtgtgca agtttctgta aatcgattaa tttttttttc tttcctcttt ttattaacct
     2101 taatttttat tttagattcc tgacttcaac tcaagacgca cagatattat aacatctgca
     2161 taataggcat ttgcaagaat tactcgtgag taaggaaaga gtgaggaact atcgcatacc
     2221 tgcatttaaa gatgccgatt tgggcgcgaa tcctttattt tggcttcacc ctcatactat
     2281 tatcagggcc agaaaaagga agtgtttccc tccttcttga attgatgtta ccctcataaa
     2341 gcacgtggcc tcttatcgag aaagaaatta ccgtcgctcg tgatttgttt gcaaaaagaa
     2401 caaaactgaa aaaacccaga cacgctcgac ttcctgtctt cctattgatt gcagcttcca
     2461 atttcgtcac acaacaaggt cctagcgacg gctcacaggt tttgtaacaa gcaatcgaag
     2521 gttctggaat ggcgggaaag ggtttagtac cacatgctat gatgcccact gtgatctcca
     2581 gagcaaagtt cgttcgatcg tactgttact ctctctcttt caaacagaat tgtccgaatc
     2641 gtgtgacaac aacagcctgt tctcacacac tcttttcttc taaccaaggg ggtggtttag
     2701 tttagtagaa cctcgtgaaa cttacattta catatatata aacttgcata aattggtcaa
     2761 tgcaagaaat acatatttgg tcttttctaa ttcgtagttt ttcaagttct tagatgcttt
     2821 ctttttctct tttttacaga tcatcaagga agtaattatc tactttttac aacaaatata
     2881 aaacaacctg cactaaaaca atggccaatt ttaatcaaag tgggaatatt gctgatagct
     2941 cattgtcctt cactttcact aacagtagca acggtccgaa cctcataaca actcaaacaa
     3001 attctcaagc gctttcacaa ccaattgcct cctctaacgt tcatgataac tttatgaata
     3061 atgaaatcac ggctagtaaa attgatgatg gtaataattc aaaaccactg tcacctggtt
     3121 ggacggacca aactgcgtat aacgcgtttg gaatcactac agagatgttt aataccacta
     3181 caatggatga tgtatataac tatctattcg atgatgaaga taccccacca aacccaaaaa
     3241 aagagccagg tccaccaaaa aagaagagaa aggttgaaaa catcaacgct gatgacacct
     3301 acagaatcat caacaagatt aaggcttgca ggtctaacaa cgatattaac cagtgtttgt
     3361 ccgacatgac caagatggtt cattgcgaat attacttgtt ggccatcatc tacccacact
     3421 ctatggttga gtccgacatt tccattttgg acaactaccc aaaaaaatgg cgtcagtatt
     3481 acgatgatgc caacttgatt aagtacgacc caatcgttga ttactcctac tctaatcatt
     3541 ctcccatcaa ctggaacatc ttcgaaaaca atgccgtcaa caaaaagtcc ccaaacgtta
     3601 tcaaagaagc taagacctat ggtttggtta ccggtttttc tttcccaatt cataccgcta
     3661 acaatggttt cggtatgttg tcttttgccc actctgaaaa ggataactac atcgactcat
     3721 tattcttgca tgcctgcatg aacattccat tgatagttcc atctctggtc gataactata
     3781 ggaagattaa cattgccaac aacaagtcca acaacgattt gaccaaaagg gaaaaagaat
     3841 gtttggcttg ggcttgcgaa ggtaaatctt ctagggatat ttccaagatt ttgggttgct
     3901 ctgaaagaac cgttactttc catttgacta acgcccaaat gaagttgaac actaccaaca
     3961 gatgccagtc tatttctaag gctattttga ccggtgctat tgattgtcca tactttaaga
     4021 attaaaggac agatgtagat acgttgttga cacttctaaa taagcgaatt tcttatgatt
     4081 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     4141 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     4201 gctttctcag gtatagcatg aggtcgctca ccgcgttatc gcgtacccaa ttcgccctat
     4261 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     4321 cctggcgtta cccctgcagg actagtgctg aggcattaat acatgggaag attcgctttt
     4381 tttttttgaa ttacaatagt atgtctgatg tctgcaagaa gtaacaggcg tgtgcacaag
     4441 aatacgtgtg tgtgcgtaag cgtatgcact ggtggcataa cttatctaag aagtatatat
     4501 cactgacata gaaatgtaga tatacaggta tttttctcga taatcgataa aaatctcgtc
     4561 gcgctgaacc aaacttggtg gttacggaga gtttttctct catcattact gtctttcgca
     4621 ttgatttccc ctttgaccga taaaatccct tggattcata agattaaaca aagaggtgat
     4681 caaagagaac cctgtgaaag tttatgttta taaccgggca taaagtgaac tagacacttt
     4741 caagaagcca accaaagcat gagtaacgaa gcttaccagc atgatcatac cgtaaatcct
     4801 caccagatta agacctcagc gcggccgcaa atttaaataa aatgaagtga agttcctata
     4861 ctttctagag aataggaact tctatagtga gtcgaataag ggcgacacaa aatttattct
     4921 aaatgcataa taaatactga taacatctta tagtttgtat tatattttgt attatcgttg
     4981 acatgtataa ttttgatatc aaaaactgat tttcccttta ttattttcga gatttatttt
     5041 cttaattctc tttaacaaac tagaaatatt gtatatacaa aaaatcataa ataatagatg
     5101 aatagtttaa ttataggtgt tcatcaatcg aaaaagcaac gtatcttatt taaagtgcgt
     5161 tgcttttttc tcatttataa ggttaaataa ttctcatata tcaagcaaag tgacaggcgc
     5221 ccttaaatat tctgacaaat gctctttccc taaactcccc ccataaaaaa acccgccgaa
     5281 gcgggttttt acgttatttg cggattaacg attactcgtt atcagaaccg cccagggggc
     5341 ccgagcttaa gactggccgt cgttttacaa cacagaaaga gtttgtagaa acgcaaaaag
     5401 gccatccgtc aggggccttc tgcttagttt gatgcctggc agttccctac tctcgccttc
     5461 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5521 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5581 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5641 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5701 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5761 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5821 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5881 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5941 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6001 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtgggctaa
     6061 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6121 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6181 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6241 cttttctacg gggtctgacg ctcagtggaa cgacgcgcgc gtaactcacg ttaagggatt
     6301 ttggtcatga gcttgcgccg tcccgtcaag tcagcgtaat gctctgctt
//
