LOCUS       sequence-209113-        8176 bp    DNA     circular SYN 15-APR-2025
DEFINITION  yeast expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8176)
  AUTHORS   Worseck JM, Grossmann A, Weimann M, Hegele A, Stelzl U
  TITLE     A stringent yeast two-hybrid matrix screening approach for 
            protein-protein interaction discovery.
  JOURNAL   Methods Mol Biol. 2012;812:63-87. doi: 10.1007/978-1-61779-455-1_4.
  PUBMED    22218854
REFERENCE   2  (bases 1 to 8176)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2012"; volume: "812:63-87. doi"; pages: " 
            10.1007/978-1-61779-455-1_4"
COMMENT     Sequence Label: pBTM116-D9
FEATURES             Location/Qualifiers
     source          1..8176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..402
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             join(429..680,681..1034)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     primer_bind     972..993
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
     protein_bind    1053..1177
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1351..1453
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1454..2113
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1520..1539)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2455..2760
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTADMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2801..2925)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      2970..3157
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3025..3049)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3210..3884)
                     /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(3885..3986)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(4000..4016)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4000..4016)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4013..4035)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4024..4040
                     /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(4048..4054,4055..4072,4073..4078))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4093..4114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4231..4248)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4402..4990)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4482..4501)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        complement(5270..5374)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5487..5492,5493..5509,5510..5515)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             5563..6753
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(5915..5934)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(6850..6872)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6972..6991
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      7008..8172
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gcatgcaact tcttttcttt ttttttcttt tctctctccc ccgttgttgt ctcaccatat
       61 ccgcaatgac aaaaaaatga tggaagacac taaaggaaaa aattaacgac aaagacagca
      121 ccaacagatg tcgttgttcc agagctgatg aggggtatct cgaagcacac gaaacttttt
      181 ccttccttca ttcacgcaca ctactctcta atgagcaacg gtatacggcc ttccttccag
      241 ttacttgaat ttgaaataaa aaaaagtttg ctgtcttgct atcaagtata aatagacctg
      301 caattattaa tcttttgttt cctcgtcatt gttctcgttc cctttcttcc ttgtttcttt
      361 ttctgcacaa tatttcaagc tataccaagc atacaatcaa ctccaagctt gaattaattc
      421 cgggcggaat gaaagcgtta acggccaggc aacaagaggt gtttgatctc atccgtgatc
      481 acatcagcca gacaggtatg ccgccgacgc gtgcggaaat cgcgcagcgt ttggggttcc
      541 gttccccaaa cgcggctgaa gaacatctga aggcgctggc acgcaaaggc gttattgaaa
      601 ttgtttccgg cgcatcacgc gggattcgtc tgttgcagga agaggaagaa gggttgccgc
      661 tggtaggtcg tgtggctgcc ggtgaaccac ttctggcgca acagcatatt gaaggtcatt
      721 atcaggtcga tccttcctta ttcaagccga atgctgattt cctgctgcgc gtcagcggga
      781 tgtcgatgaa agatatcggc attatggatg gtgacttgct ggcagtgcat aaaactcagg
      841 atgtacgtaa cggtcaggtc gttgtcgcac gtattgatga cgaggttacc gttaagcgcc
      901 tgaaaaaaca gggcaataaa gtcgaactgt tgccagaaaa tagcgagttt aaaccaattg
      961 tcgtagatct tcgtcagcag agcttcacca ttgaagggct ggcggttggg gttattcgca
     1021 acggcgactg gctggaattc atgtcgaatc aaacaagttt gtacaaaaaa gctgaacgag
     1081 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
     1141 cataatactg taaaacacaa catatccagt cactatggcg gccgcgggtg atgctgccaa
     1201 cttagcggcc gctaagttgg cagcatcacc cgacgcactt tgcgccgaat aaatacctgt
     1261 gacggaagat cacttcgcag aataaataaa tcctggtgtc cctgttgata ccgggaagcc
     1321 ctgggccaac ttttggcgaa aatgagacgt tgatcggcac gtaagaggtt ccaactttca
     1381 ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc
     1441 taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
     1501 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
     1561 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
     1621 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc
     1681 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
     1741 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
     1801 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     1861 agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt
     1921 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata
     1981 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg
     2041 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
     2101 gggcggggcg taatctagag gatccggctt actaaaagcc agataacagt atgcgtattt
     2161 gcgcgctgat tttcgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa
     2221 aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg
     2281 ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag
     2341 cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg
     2401 cccggtttat tgaaatgaac ggctcttttg ctgacgagaa caggggctgg tgaaatgcag
     2461 tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt
     2521 gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg
     2581 tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc
     2641 atgatgaccg ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat
     2701 ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa
     2761 atgtcaggct cccttataca cagccagtct gcaggtcgac catagtgact ggatatgttg
     2821 tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt
     2881 tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggttcgat tcgacggtac
     2941 cgcggccgct gcagccaagc taattccggg cgaatttctt atgatttatg atttttatta
     3001 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
     3061 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     3121 agcatgaggt cgctcttatt gaccacacct ctaccggcat gccggcaagt gcacaaacaa
     3181 tacttaaata aatactactc agtaataacc tatttcttag catttttgac gaaatttgct
     3241 attttgttag agtcttttac accatttgtc tccacacctc cgcttacatc aacaccaata
     3301 acgccattta atctaagcgc atcaccaaca ttttctggcg tcagtccacc agctaacata
     3361 aaatgtaagc tttcggggct ctcttgcctt ccaacccagt cagaaatcga gttccaatcc
     3421 aaaagttcac ctgtcccacc tgcttctgaa tcaaacaagg gaataaacga atgaggtttc
     3481 tgtgaagctg cactgagtag tatgttgcag tcttttggaa atacgagtct tttaataact
     3541 ggcaaaccga ggaactcttg gtattcttgc cacgactcat ctccatgcag ttggacgata
     3601 tcaatgccgt aatcattgac cagagccaaa acatcctcct taggttgatt acgaaacacg
     3661 ccaaccaagt atttcggagt gcctgaacta tttttatatg cttttacaag acttgaaatt
     3721 ttccttgcaa taaccgggtc aattgttctc tttctattgg gcacacatat aatacccagc
     3781 aagtcagcat cggaatctag agcacattct gcggcctctg tgctctgcaa gccgcaaact
     3841 ttcaccaatg gaccagaact acctgtgaaa ttaataacag acatactcca agctgccttt
     3901 gtgtgcttaa tcacgtatac tcacgtgctc aatagtcacc aatgccctcc ctcttggccc
     3961 tctccttttc ttttttcgac cgaattaatt cgtaatcatg tcatagctgt ttcctgtgtg
     4021 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4081 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4141 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4201 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4261 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4321 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4381 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4441 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4501 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4561 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4621 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4681 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4741 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4801 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4861 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4921 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4981 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5041 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5101 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5161 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5221 agttgcctga ctcgggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5281 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5341 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5401 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     5461 cctttcgtct tcaagaattc tcatgtttga cagcttatca tcgataagct ttaatgcggt
     5521 agtttatcac agttaaattg ctaacgcagt caggcaccgt gtatgaaatc taacaatgcg
     5581 ctcatcgtca tcctcggcac cgtcaccctg gatgctgtag gcataggctt ggttatgccg
     5641 gtactgccgg gcctcttgcg ggatatcgtc cattccgaca gcatcgccag tcactatggc
     5701 gtgctgctag cgctatatgc gttgatgcaa tttctatgcg cacccgttct cggagcactg
     5761 tccgaccgct ttggccgccg cccagtcctg ctcgcttcgc tacttggagc cactatcgac
     5821 tacgcgatca tggcgaccac acccgtcctg tggatcctct acgccggacg catcgtggcc
     5881 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
     5941 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
     6001 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
     6061 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
     6121 ggagagcgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg
     6181 cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga
     6241 caggtgccgg cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg
     6301 atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc
     6361 actggtcccg ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc
     6421 gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag gctggatggc cttccccatt
     6481 atgattcttc tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg
     6541 caggtagatg acgaccatca gggacagctt caaggatcgc tcgcggctct taccagccta
     6601 acttcgatca ttggaccgct gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg
     6661 aacgggttgg catggattgt aggcgccgcc ctataccttg tctgcctccc cgcgttgcgt
     6721 cgcggtgcat ggagccgggc cacctcgacc tgacccctaa gaaaccatta ttatcatgac
     6781 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     6841 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     6901 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     6961 gcttaactat gcggcatcag agcagattgt actgagagtg caccataacg catttaagca
     7021 taaacacgca ctatgccgtt cttctcatgt atatatatat acaggcaaca cgcagatata
     7081 ggtgcgacgt gaacagtgag ctgtatgtgc gcagctcgcg ttgcattttc ggaagcgctc
     7141 gttttcggaa acgctttgaa gttcctattc cgaagttcct attctctagc tagaaagtat
     7201 aggaacttca gagcgctttt gaaaaccaaa agcgctctga agacgcactt tcaaaaaacc
     7261 aaaaacgcac cggactgtaa cgagctacta aaatattgcg aataccgctt ccacaaacat
     7321 tgctcaaaag tatctctttg ctatatatct ctgtgctata tccctatata acctacccat
     7381 ccacctttcg ctccttgaac ttgcatctaa actcgacctc tacatttttt atgtttatct
     7441 ctagtattac tctttagaca aaaaaattgt agtaagaact attcatagag tgaatcgaaa
     7501 acaatacgaa aatgtaaaca tttcctatac gtagtatata gagacaaaat agaagaaacc
     7561 gttcataatt ttctgaccaa tgaagaatca tcaacgctat cactttctgt tcacaaagta
     7621 tgcgcaatcc acatcggtat agaatataat cggggatgcc tttatcttga aaaaatgcac
     7681 ccgcagcttc gctagtaatc agtaaacgcg ggaagtggag tcaggctttt tttatggaag
     7741 agaaaataga caccaaagta gccttcttct aaccttaacg gacctacagt gcaaaaagtt
     7801 atcaagagac tgcattatag agcgcacaaa ggagaaaaaa agtaatctaa gatgctttgt
     7861 tagaaaaata gcgctctcgg gatgcatttt tgtagaacaa aaaagaagta tagattcttt
     7921 gttggtaaaa tagcgctctc gcgttgcatt tctgttctgt aaaaatgcag ctcagattct
     7981 ttgtttgaaa aattagcgct ctcgcgttgc atttttgttt tacaaaaatg aagcacagat
     8041 tcttcgttgg taaaatagcg ctttcgcgtt gcatttctgt tctgtaaaaa tgcagctcag
     8101 attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttctacaa aatgaagcac
     8161 agatgcttcg ttgctt
//
