LOCUS       sequence-484559-        7048 bp    DNA     circular SYN 25-AUG-2025
DEFINITION  Yeast two hybrid vector expressing BD-PYR1_star-MANDI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7048)
  AUTHORS   Park SY, Qiu J, Wei S, Peterson FC, Beltran J, Medina-Cucurella AV, 
            Vaidya AS, Xing Z, Volkman BF, Nusinow DA, Whitehead TA, Wheeldon I,
            Cutler SR
  TITLE     An orthogonalized PYR1-based CID module with reprogrammable 
            ligand-binding specificity.
  JOURNAL   Nat Chem Biol. 2024 Jan;20(1):103-110. doi: 
            10.1038/s41589-023-01447-7. Epub 2023 Oct 23.
  PUBMED    37872402
REFERENCE   2  (bases 1 to 7048)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41589-023-01447-7"; journalName: "Nat Chem Biol"; date: 
            "2024-01"; volume: "20"; issue: "1"; pages: "103-110"
COMMENT     Sequence Label: pBD-PYR1_star-MANDI
FEATURES             Location/Qualifiers
     source          1..7048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..406
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             434..874
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     primer_bind     complement(1473..1492)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1474..1492)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1514..1701
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1569..1593)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(1754..2428)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(2429..2530)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2544..2560)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2544..2560)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2557..2579)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2568..2584
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2592..2598,2599..2616,2617..2622))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2637..2658
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        complement(2730..2834)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2861..3316
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2948..2967)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3158..3179
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3411..3428)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3582..4170)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3662..3681)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4627..4729
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4730..5389
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4796..4815)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     5670..5688
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5726..5748)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5848..5867
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      5884..7048
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc
       61 atatccgcaa tgacaaaaaa atgatggaag acactaaagg aaaaaattaa cgacaaagac
      121 agcaccaaca gatgtcgttg ttccagagct gatgaggggt atctcgaagc acacgaaact
      181 ttttccttcc ttcattcacg cacactactc tctaatgagc aacggtatac ggccttcctt
      241 ccagttactt gaatttgaaa taaaaaaaag tttgctgtct tgctatcaag tataaataga
      301 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
      361 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gcttgaagca
      421 agcctcctga aagatgaagc tactgtcttc tatcgaacaa gcatgcgata tttgccgact
      481 taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg
      541 ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac atctgacaga
      601 agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc ctcgagaaga
      661 ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt taacaggatt
      721 atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt cagtggagac
      781 tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat cggaagagag
      841 tagtaacaaa ggtcaaagac agttgactgt atcgccggaa ttcatgcctt cggagttaac
      901 accagaagaa cgatcggaac taaaaaactc aatcgccgag ttccacacat accaactcga
      961 tccaggaagc tgttcatcac tccacgcgca acgaatccac gcgcctccgg aactcgtctg
     1021 gtcaatcgta cgacgattcg acaaaccaca aacacaccgc cacttcatca aatcctgctc
     1081 cgtcgaacaa aacttcgaga tgcgcgtcgg atgcacgcgc gacatcatcg tcatcagtgg
     1141 attaccggcg aacacatcaa cggaaagact cgatatactc gacgacgaac ggagagttac
     1201 cggagccagt atcatcggag gcgaacatag gctgacgaat tacaaaggcg ttacgacggt
     1261 gcatcggttc gagaaagaga atcggatctg gacggtggtt ttggaatctt acgtcgttga
     1321 tatgccggaa ggtaactcgg aggatgatac tcgtattgtt gtggatgacg ttgtgaagct
     1381 taatttgcag aaactcgcga cggttgctga agctatggct cgtaactccg gtgacggaag
     1441 tggttctcag gtgacgtgag tcgactctag agccctatag tgagtcgtat tacctcagcc
     1501 aagctaattc cgggcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
     1561 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt
     1621 cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagcatg aggtcgctct
     1681 tattgaccac acctctaccg gcatgccggc aagtgcacaa acaatactta aataaatact
     1741 actcagtaat aacctatttc ttagcatttt tgacgaaatt tgctattttg ttagagtctt
     1801 ttacaccatt tgtctccaca cctccgctta catcaacacc aataacgcca tttaatctaa
     1861 gcgcatcacc aacattttct ggcgtcagtc caccagctaa cataaaatgt aagctttcgg
     1921 ggctctcttg ccttccaacc cagtcagaaa tcgagttcca atccaaaagt tcacctgtcc
     1981 cacctgcttc tgaatcaaac aagggaataa acgaatgagg tttctgtgaa gctgcactga
     2041 gtagtatgtt gcagtctttt ggaaatacga gtcttttaat aactggcaaa ccgaggaact
     2101 cttggtattc ttgccacgac tcatctccat gcagttggac gatatcaatg ccgtaatcat
     2161 tgaccagagc caaaacatcc tccttaggtt gattacgaaa cacgccaacc aagtatttcg
     2221 gagtgcctga actattttta tatgctttta caagacttga aattttcctt gcaataaccg
     2281 ggtcaattgt tctctttcta ttgggcacac atataatacc cagcaagtca gcatcggaat
     2341 ctagagcaca ttctgcggcc tctgtgctct gcaagccgca aactttcacc aatggaccag
     2401 aactacctgt gaaattaata acagacatac tccaagctgc ctttgtgtgc ttaatcacgt
     2461 atactcacgt gctcaatagt caccaatgcc ctccctcttg gccctctcct tttctttttt
     2521 cgaccgaatt aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2581 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2641 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2701 tcgtgccagc tggttctttc cgcctcagga ctcttccttt ttcaatatta ttgaagcatt
     2761 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     2821 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     2881 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2941 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3001 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3061 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3121 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3181 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3241 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3301 ttaacgctta caatttacgc gcctaaatta tgcggccgca tataactgca ttaatgaatc
     3361 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3421 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3481 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3541 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3601 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3661 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3721 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3781 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3841 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3901 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3961 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4021 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4081 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4141 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4201 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4261 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4321 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4381 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4441 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4501 ccagatttat cagcaataaa ccagccagcc gcctgtgacg gaagatcact tcgcagaata
     4561 aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg
     4621 agacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
     4681 gggcgtattt tttgagttgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     4741 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     4801 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     4861 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     4921 ccgcctgatg aatgctcatc cggaattacg tatggcaatg aaagacggtg agctggtgat
     4981 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     5041 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     5101 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     5161 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     5221 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     5281 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     5341 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg
     5401 cagttattgg tgcccttaaa cgcctggttg ctacgcctga ataagtgata ataagcggat
     5461 gaatggcaga aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa
     5521 atagccgctt atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc
     5581 gtgtgcttct caaatgcctg aggccagttt gctcaggctc tccccgtgga ggctaagaaa
     5641 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     5701 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     5761 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     5821 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5881 ataacgcatt taagcataaa cacgcactat gccgttcttc tcatgtatat atatatacag
     5941 gcaacacgca gatataggtg cgacgtgaac agtgagctgt atgtgcgcag ctcgcgttgc
     6001 attttcggaa gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa gttcctattc
     6061 tctagctaga aagtatagga acttcagagc gcttttgaaa accaaaagcg ctctgaagac
     6121 gcactttcaa aaaaccaaaa acgcaccgga ctgtaacgag ctactaaaat attgcgaata
     6181 ccgcttccac aaacattgct caaaagtatc tctttgctat atatctctgt gctatatccc
     6241 tatataacct acccatccac ctttcgctcc ttgaacttgc atctaaactc gacctctaca
     6301 ttttttatgt ttatctctag tattactctt tagacaaaaa aattgtagta agaactattc
     6361 atagagtgaa tcgaaaacaa tacgaaaatg taaacatttc ctatacgtag tatatagaga
     6421 caaaatagaa gaaaccgttc ataattttct gaccaatgaa gaatcatcaa cgctatcact
     6481 ttctgttcac aaagtatgcg caatccacat cggtatagaa tataatcggg gatgccttta
     6541 tcttgaaaaa atgcacccgc agcttcgcta gtaatcagta aacgcgggaa gtggagtcag
     6601 gcttttttta tggaagagaa aatagacacc aaagtagcct tcttctaacc ttaacggacc
     6661 tacagtgcaa aaagttatca agagactgca ttatagagcg cacaaaggag aaaaaaagta
     6721 atctaagatg ctttgttaga aaaatagcgc tctcgggatg catttttgta gaacaaaaaa
     6781 gaagtataga ttctttgttg gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa
     6841 atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca
     6901 aaaatgaagc acagattctt cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg
     6961 taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt
     7021 ctacaaaatg aagcacagat gcttcgtt
//
