LOCUS       sequence-484552-        7049 bp    DNA     circular SYN 25-AUG-2025
DEFINITION  Yeast two hybrid vector expressing BD-PYR1 (wild type).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7049)
  AUTHORS   Park SY, Fung P, Nishimura N, Jensen DR, Fujii H, Zhao Y, Lumba S, 
            Santiago J, Rodrigues A, Chow TF, Alfred SE, Bonetta D, Finkelstein 
            R, Provart NJ, Desveaux D, Rodriguez PL, McCourt P, Zhu JK, 
            Schroeder JI, Volkman BF, Cutler SR
  TITLE     Abscisic acid inhibits type 2C protein phosphatases via the PYR/PYL 
            family of START proteins.
  JOURNAL   Science. 2009 May 22;324(5930):1068-71. doi: 
            10.1126/science.1173041. Epub 2009 Apr 30.
  PUBMED    19407142
REFERENCE   2  (bases 1 to 7049)
  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: 
            "10.1126/science.1173041"; journalName: "Science"; date: 
            "2009-05-22- 22"; volume: "324"; issue: "5930"; pages: "1068-71"
COMMENT     Sequence Label: pBD-PYR1
FEATURES             Location/Qualifiers
     source          1..7049
                     /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(1474..1493)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1475..1493)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1515..1702
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1570..1594)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(1755..2429)
                     /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(2430..2531)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2545..2561)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2545..2561)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2558..2580)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2569..2585
                     /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(2593..2599,2600..2617,2618..2623))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2638..2659
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        complement(2731..2835)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2862..3317
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2949..2968)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3159..3180
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3412..3429)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3583..4171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3663..3682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4628..4730
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4731..5390
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4797..4816)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     5671..5689
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5727..5749)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5849..5868
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      5885..7049
                     /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 gtccccgctg 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 aacatacaaa cacttcatca aatcctgctc
     1081 cgtcgaacaa aacttcgaga tgcgcgtcgg atgcacgcgc gacgtgatcg tcatcagtgg
     1141 attaccggcg aacacatcaa cggaaagact cgatatactc gacgacgaac ggagagttac
     1201 cggattcagt atcatcggag gcgaacatag gctgacgaat tacaaatccg ttacgacggt
     1261 gcatcggttc gagaaagaga atcggatctg gacggtggtt ttggaatctt acgtcgttga
     1321 tatgccggaa ggtaactcgg aggatgatac tcgtatgttt gctgatacgg ttgtgaagct
     1381 taatttgcag aaactcgcga cggttgctga agctatggct cgtaactccg gtgacggaag
     1441 tggttctcag gtgacgtgag gtcgactcta gagccctata gtgagtcgta ttactgcagc
     1501 caagctaatt ccgggcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     1561 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
     1621 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat gaggtcgctc
     1681 ttattgacca cacctctacc ggcatgccgg caagtgcaca aacaatactt aaataaatac
     1741 tactcagtaa taacctattt cttagcattt ttgacgaaat ttgctatttt gttagagtct
     1801 tttacaccat ttgtctccac acctccgctt acatcaacac caataacgcc atttaatcta
     1861 agcgcatcac caacattttc tggcgtcagt ccaccagcta acataaaatg taagctttcg
     1921 gggctctctt gccttccaac ccagtcagaa atcgagttcc aatccaaaag ttcacctgtc
     1981 ccacctgctt ctgaatcaaa caagggaata aacgaatgag gtttctgtga agctgcactg
     2041 agtagtatgt tgcagtcttt tggaaatacg agtcttttaa taactggcaa accgaggaac
     2101 tcttggtatt cttgccacga ctcatctcca tgcagttgga cgatatcaat gccgtaatca
     2161 ttgaccagag ccaaaacatc ctccttaggt tgattacgaa acacgccaac caagtatttc
     2221 ggagtgcctg aactattttt atatgctttt acaagacttg aaattttcct tgcaataacc
     2281 gggtcaattg ttctctttct attgggcaca catataatac ccagcaagtc agcatcggaa
     2341 tctagagcac attctgcggc ctctgtgctc tgcaagccgc aaactttcac caatggacca
     2401 gaactacctg tgaaattaat aacagacata ctccaagctg cctttgtgtg cttaatcacg
     2461 tatactcacg tgctcaatag tcaccaatgc cctccctctt ggccctctcc ttttcttttt
     2521 tcgaccgaat taattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct
     2581 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2641 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2701 gtcgtgccag ctggttcttt ccgcctcagg actcttcctt tttcaatatt attgaagcat
     2761 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2821 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     2881 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2941 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3001 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3061 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3121 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3181 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3241 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3301 attaacgctt acaatttacg cgcctaaatt atgcggccgc atataactgc attaatgaat
     3361 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3421 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3481 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3541 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3601 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3661 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3721 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3781 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3841 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3901 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3961 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4021 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4081 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4141 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4201 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4261 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4321 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4381 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4441 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgggacccac gctcaccggc
     4501 tccagattta tcagcaataa accagccagc cgcctgtgac ggaagatcac ttcgcagaat
     4561 aaataaatcc tggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat
     4621 gagacgttga tcggcacgta agaggttcca actttcacca taatgaaata agatcactac
     4681 cgggcgtatt ttttgagttg tcgagatttt caggagctaa ggaagctaaa atggagaaaa
     4741 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     4801 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
     4861 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     4921 cccgcctgat gaatgctcat ccggaattac gtatggcaat gaaagacggt gagctggtga
     4981 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     5041 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     5101 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     5161 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     5221 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
     5281 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     5341 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
     5401 gcagttattg gtgcccttaa acgcctggtt gctacgcctg aataagtgat aataagcgga
     5461 tgaatggcag aaattcgaaa gcaaattcga cccggtcgtc ggttcagggc agggtcgtta
     5521 aatagccgct tatgtctatt gctggtttac cggtttattg actaccggaa gcagtgtgac
     5581 cgtgtgcttc tcaaatgcct gaggccagtt tgctcaggct ctccccgtgg aggctaagaa
     5641 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     5701 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     5761 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     5821 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     5881 cataacgcat ttaagcataa acacgcacta tgccgttctt ctcatgtata tatatataca
     5941 ggcaacacgc agatataggt gcgacgtgaa cagtgagctg tatgtgcgca gctcgcgttg
     6001 cattttcgga agcgctcgtt ttcggaaacg ctttgaagtt cctattccga agttcctatt
     6061 ctctagctag aaagtatagg aacttcagag cgcttttgaa aaccaaaagc gctctgaaga
     6121 cgcactttca aaaaaccaaa aacgcaccgg actgtaacga gctactaaaa tattgcgaat
     6181 accgcttcca caaacattgc tcaaaagtat ctctttgcta tatatctctg tgctatatcc
     6241 ctatataacc tacccatcca cctttcgctc cttgaacttg catctaaact cgacctctac
     6301 attttttatg tttatctcta gtattactct ttagacaaaa aaattgtagt aagaactatt
     6361 catagagtga atcgaaaaca atacgaaaat gtaaacattt cctatacgta gtatatagag
     6421 acaaaataga agaaaccgtt cataattttc tgaccaatga agaatcatca acgctatcac
     6481 tttctgttca caaagtatgc gcaatccaca tcggtataga atataatcgg ggatgccttt
     6541 atcttgaaaa aatgcacccg cagcttcgct agtaatcagt aaacgcggga agtggagtca
     6601 ggcttttttt atggaagaga aaatagacac caaagtagcc ttcttctaac cttaacggac
     6661 ctacagtgca aaaagttatc aagagactgc attatagagc gcacaaagga gaaaaaaagt
     6721 aatctaagat gctttgttag aaaaatagcg ctctcgggat gcatttttgt agaacaaaaa
     6781 agaagtatag attctttgtt ggtaaaatag cgctctcgcg ttgcatttct gttctgtaaa
     6841 aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttttac
     6901 aaaaatgaag cacagattct tcgttggtaa aatagcgctt tcgcgttgca tttctgttct
     6961 gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt
     7021 tctacaaaat gaagcacaga tgcttcgtt
//
