LOCUS       sequence-368327-        7671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Y2H vector compatible with pJS647 PYR1 insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7671)
  AUTHORS   Beltran J, Steiner PJ, Bedewitz M, Wei S, Peterson FC, Li Z, Hughes 
            BE, Hartley Z, Robertson NR, Medina-Cucurella AV, Baumer ZT, Leonard
            AC, Park SY, Volkman BF, Nusinow DA, Zhong W, Wheeldon I, Cutler SR,
            Whitehead TA
  TITLE     Rapid biosensor development using plant hormone receptors as 
            reprogrammable scaffolds.
  JOURNAL   Nat Biotechnol. 2022 Jun 20. pii: 10.1038/s41587-022-01364-5. doi: 
            10.1038/s41587-022-01364-5.
  PUBMED    35726092
REFERENCE   2  (bases 1 to 7671)
  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: "Nat 
            Biotechnol. 2022 Jun 20. pii: 10.1038/s41587-022-01364-5. doi: 
            10.1038/s41587-022-01364-5."
COMMENT     Sequence Label: pJS646
FEATURES             Location/Qualifiers
     source          1..7671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..113
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    185..206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(221..226,227..244,245..251)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    259..275
                     /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)."
     primer_bind     264..286
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     283..299
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     283..299
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        313..414
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             415..1089
                     /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"
     terminator      complement(1142..1329)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     1250..1274
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     1351..1370
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1351..1369
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(1433..1460)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1476..1547)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1559..2272)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     RBS             2279..2290
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(2592..3032)
                     /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"
     promoter        complement(3060..3461)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     rep_origin      3466..4630
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     complement(4647..4666)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4766..4788
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4826..4844)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(5125..5784)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5699..5718
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5785..5887)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      6344..6932
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6833..6852
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7086..7103
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7198..7653)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7335..7356)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7547..7566
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
       61 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
      121 cggaaagaac cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
      181 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
      241 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca
      301 tgattacgaa ttaattcggt cgaaaaaaga aaaggagagg gccaagaggg agggcattgg
      361 tgactattga gcacgtgagt atacgtgatt aagcacacaa aggcagcttg gagtatgtct
      421 gttattaatt tcacaggtag ttctggtcca ttggtgaaag tttgcggctt gcagagcaca
      481 gaggccgcag aatgtgctct agattccgat gctgacttgc tgggtattat atgtgtgccc
      541 aatagaaaga gaacaattga cccggttatt gcaaggaaaa tttcaagtct tgtaaaagca
      601 tataaaaata gttcaggcac tccgaaatac ttggttggcg tgtttcgtaa tcaacctaag
      661 gaggatgttt tggctctggt caatgattac ggcattgata tcgtccaact gcatggagat
      721 gagtcgtggc aagaatacca agagttcctc ggtttgccag ttattaaaag actcgtattt
      781 ccaaaagact gcaacatact actcagtgca gcttcacaga aacctcattc gtttattccc
      841 ttgtttgatt cagaagcagg tgggacaggt gaacttttgg attggaactc gatttctgac
      901 tgggttggaa ggcaagagag ccccgaaagc ttacatttta tgttagctgg tggactgacg
      961 ccagaaaatg ttggtgatgc gcttagatta aatggcgtta ttggtgttga tgtaagcgga
     1021 ggtgtggaga caaatggtgt aaaagactct aacaaaatag caaatttcgt caaaaatgct
     1081 aagaaatagg ttattactga gtagtattta tttaagtatt gtttgtgcac ttgccggcat
     1141 gccggtagag gtgtggtcaa taagagcgac ctcatgctat acctgagaaa gcaacctgac
     1201 ctacaggaaa gagttactca agaataagaa ttttcgtttt aaaacctaag agtcacttta
     1261 aaatttgtat acacttattt tttttataac ttatttaata ataaaaatca taaatcataa
     1321 gaaattcgcc cggaattagc ttggctgagg taatacgact cactataggg ctctagagtc
     1381 gaccgtcacc tgagaaccac ttccgtcacc ggagttacgt gagacctata aacgcagaaa
     1441 ggcccacccg aaggtgagcc agtgtgactc tagtagagag cgttcaccga caaacaacag
     1501 ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctggtcatt
     1561 tgtacagttc atccatacca tgcgtgatgc ccgctgcggt tacgaactcc agcagaacca
     1621 tatgatcgcg tttctcgttc ggatctttag acagaacgct ttgcgtgctc agatagtgat
     1681 tgtctggcag cagaacagga ccatcaccga ttggagtgtt ttgctggtag tgatcagcca
     1741 gctgcacgct gccatcctcc acgttgtggc gaattttaaa attcgcttta atgccatttt
     1801 tttgtttatc ggcggtgatg taaacattgt ggctgttaaa attgtattcc agcttatggc
     1861 ccaggatatt gccgtcctct ttaaagtcaa tgcctttcag ctcaatgcgg tttaccaggg
     1921 tatcgccttc aaatttcact tccgcacgcg ttttgtacgt gccgtcatcc ttaaaggaaa
     1981 tcgtgcgttc ctgcacatag ccttccggca tggcggactt gaagaagtca tgctgcttca
     2041 tatggtccgg ataacgagca aagcactgaa caccataagt cagcgtcgtt accagagtcg
     2101 gccaaggaac cggcagttta ccagtagtac agatgaactt cagcgtcagt ttaccattag
     2161 ttgcgtcacc ttcaccctcg ccacgcacgg aaaacttatg accgttgaca tcaccatcca
     2221 gttccaccag aatagggacg acaccagtga acagctcttc gcctttacgc atctagtatt
     2281 tctcctcttt ctctagtagc taagatccga acctgccgtt tcttgagttg ccgcgatgtt
     2341 aagaaaacat tcataaatta aatgtgaatt gccgcacaca ttattaaata agatttacaa
     2401 aatgttcaaa atgacgcatg aaatcacgtt tcactttcgg tctcacagac gagttccgga
     2461 ggcgcgtgga ttcgttgcgc gtggagtgat gaacagcttc ctggatcgag ttggtatgtg
     2521 tggaactcgg cgattgagtt ttttagttcc gatcgttctt ctggtgttaa ctccgaaggc
     2581 atgaattccg gcgatacagt caactgtctt tgacctttgt tactactctc ttccgatgat
     2641 gatgtcgcac ttattctatg ctgtctcaat gttagaggca tatcagtctc cactgaagcc
     2701 aatctatctg tgacggcatc tttattcaca ttatcttgta caaataatcc tgttaacaat
     2761 gcttttatat cctgtaaaga atccattttc aaaatcatgt caaggtcttc tcgaggaaaa
     2821 atcagtagaa atagctgttc cagtctttct agccttgatt ccacttctgt cagatgtgcc
     2881 ctagtcagcg gagatctttt ggttttggga gagtagcgac actcccagtt gttcttcaga
     2941 cacttggcgc acttcggttt ttctttggag cacttgagct ttttaagtcg gcaaatatcg
     3001 catgcttgtt cgatagaaga cagtagcttc atctttcagg aggcttgctt caagcttgga
     3061 gttgattgta tgcttggtat agcttgaaat attgtgcaga aaaagaaaca aggaagaaag
     3121 ggaacgagaa caatgacgag gaaacaaaag attaataatt gcaggtctat ttatacttga
     3181 tagcaagaca gcaaactttt ttttatttca aattcaagta actggaagga aggccgtata
     3241 ccgttgctca ttagagagta gtgtgcgtga atgaaggaag gaaaaagttt cgtgtgcttc
     3301 gagatacccc tcatcagctc tggaacaacg acatctgttg gtgctgtctt tgtcgttaat
     3361 tttttccttt agtgtcttcc atcatttttt tgtcattgcg gatatggtga gacaacaacg
     3421 ggggagagag aaaagaaaaa aaaagaaaag aagttgcatg caagcaacga agcatctgtg
     3481 cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     3541 agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc
     3601 tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     3661 atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa
     3721 gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca
     3781 aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac
     3841 tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt
     3901 ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg
     3961 cattttttca agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac
     4021 tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt
     4081 ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt
     4141 cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga
     4201 taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg
     4261 ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg
     4321 tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt
     4381 tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa agcgctctga agttcctata
     4441 ctttctagct agagaatagg aacttcggaa taggaacttc aaagcgtttc cgaaaacgag
     4501 cgcttccgaa aatgcaacgc gagctgcgca catacagctc actgttcacg tcgcacctat
     4561 atctgcgtgt tgcctgtata tatatataca tgagaagaac ggcatagtgc gtgtttatgc
     4621 ttaaatgcgt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4681 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4741 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4801 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4861 atgataataa tggtttctta gcctccacgg ggagagcctg agcaaactgg cctcaggcat
     4921 ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac cagcaataga
     4981 cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcgaat ttgctttcga
     5041 atttctgcca ttcatccgct tattatcact tattcaggcg tagcaaccag gcgtttaagg
     5101 gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc agtactgttg
     5161 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
     5221 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     5281 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     5341 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     5401 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     5461 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     5521 aacactatcc catatcacca gctcaccgtc tttcattgcc atacgtaatt ccggatgagc
     5581 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     5641 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     5701 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     5761 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgacaactc
     5821 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     5881 ccgatcaacg tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacagggac
     5941 accaggattt atttattctg cgaagtgatc ttccgtcaca ggcggctggc tggtttattg
     6001 ctgataaatc tggagccggt gagcgtggat ctcgcggtat cattgcagca ctggggccag
     6061 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6121 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6181 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6241 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6301 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6361 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6421 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6481 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6541 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6601 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6661 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6721 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6781 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6841 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6901 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6961 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7021 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7081 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     7141 ccgcgcgttg gccgattcat taatgcagtt atatgcggcc gcataattta ggcgcgtaaa
     7201 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     7261 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     7321 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     7381 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     7441 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     7501 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     7561 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     7621 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg g
//
