LOCUS       sequence-374353-        6636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression hexamer component in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6636)
  AUTHORS   Sang D, Shu T, Pantoja CF, Ibanez de Opakua A, Zweckstetter M, Holt 
            LJ
  TITLE     Condensed-phase signaling can expand kinase specificity and respond 
            to macromolecular crowding.
  JOURNAL   Mol Cell. 2022 Oct 6;82(19):3693-3711.e10. doi: 
            10.1016/j.molcel.2022.08.016. Epub 2022 Sep 14.
  PUBMED    36108633
REFERENCE   2  (bases 1 to 6636)
  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"; doi: 
            "10.1016/j.molcel.2022.08.016"; journalName: "Mol Cell"; date: 
            "2022-10-6- 6"; volume: "82"; issue: "19"; pages: "3693-3711.e10"
COMMENT     Sequence Label: pRS304-PTDH3-HEXAMER
FEATURES             Location/Qualifiers
     source          1..6636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..40)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(194..782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(274..293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(953..1744,1745..1813))
                     /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"
     primer_bind     1576..1595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1814..1918)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1986..2004
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2042..2064)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2164..2183
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2204..2485
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             2486..3160
                     /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"
     rep_origin      complement(3263..3718)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3400..3421)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3612..3631
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3844..3866
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3858..3875
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3859..3875
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3882..3901
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3882..3900
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3929..4571
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     4465..4488
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     4577..4593
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4577..4593
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(6025..6041)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6026..6042)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      6044..6291
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6060..6078)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(6314..6332)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(6389..6407)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6428..6444)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6428..6444)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6441..6463)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6452..6468
                     /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(6476..6482,6483..6500,6501..6506))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6521..6542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
       61 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      121 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      181 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      241 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      301 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      361 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      421 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      481 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      541 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      601 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      661 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      721 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      781 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      841 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      901 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
      961 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1021 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1081 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1141 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1201 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1261 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1321 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1381 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1441 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1501 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1561 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1621 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1681 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1741 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1801 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1861 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1921 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     1981 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     2041 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     2101 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     2161 atgcggcatc agagcagatt gtactgagag tgcaccatag atcaacgaca ttactatata
     2221 tataatatag gaagcattta atagaacagc atcgtaatat atgtgtactt tgcagttatg
     2281 acgccagatg gcagtagtgg aagatattct ttattgaaaa atagcttgtc accttacgta
     2341 caatcttgat ccggagcttt tctttttttg ccgattaaga attaattcgg tcgaaaaaag
     2401 aaaaggagag ggccaagagg gagggcattg gtgactattg agcacgtgag tatacgtgat
     2461 taagcacaca aaggcagctt ggagtatgtc tgttattaat ttcacaggta gttctggtcc
     2521 attggtgaaa gtttgcggct tgcagagcac agaggccgca gaatgtgctc tagattccga
     2581 tgctgacttg ctgggtatta tatgtgtgcc caatagaaag agaacaattg acccggttat
     2641 tgcaaggaaa atttcaagtc ttgtaaaagc atataaaaat agttcaggca ctccgaaata
     2701 cttggttggc gtgtttcgta atcaacctaa ggaggatgtt ttggctctgg tcaatgatta
     2761 cggcattgat atcgtccaac tgcatggaga tgagtcgtgg caagaatacc aagagttcct
     2821 cggtttgcca gttattaaaa gactcgtatt tccaaaagac tgcaacatac tactcagtgc
     2881 agcttcacag aaacctcatt cgtttattcc cttgtttgat tcagaagcag gtgggacagg
     2941 tgaacttttg gattggaact cgatttctga ctgggttgga aggcaagaga gccccgaaag
     3001 cttacatttt atgttagctg gtggactgac gccagaaaat gttggtgatg cgcttagatt
     3061 aaatggcgtt attggtgttg atgtaagcgg aggtgtggag acaaatggtg taaaagactc
     3121 taacaaaata gcaaatttcg tcaaaaatgc taagaaatag gttattactg agtagtattt
     3181 atttaagtat tgtttgtgca cttgccgatc tatgcggtgt gaaataccgc acagatgcgt
     3241 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     3301 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3361 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3421 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3481 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3541 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3601 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3661 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     3721 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     3781 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     3841 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg
     3901 gcgaattgga gctctcagtt cgagtttatc attatcaata ctgccatttc aaagaatacg
     3961 taaataatta atagtagtga ttttcctaac tttatttagt caaaaaatta gccttttaat
     4021 tctgctgtaa cccgtacatg cccaaaatag ggggcgggtt acacagaata tataacatcg
     4081 taggtgtctg ggtgaacagt ttattcctgg catccactaa atataatgga gcccgctttt
     4141 taagctggca tccagaaaaa aaaagaatcc cagcaccaaa atattgtttt cttcaccaac
     4201 catcagttca taggtccatt ctcttagcgc aactacagag aacaggggca caaacaggca
     4261 aaaaacgggc acaacctcaa tggagtgatg caacctgcct ggagtaaatg atgacacaag
     4321 gcaattgacc cacgcatgta tctatctcat tttcttacac cttctattac cttctgctct
     4381 ctctgatttg gaaaaagctg aaaaaaaagg ttgaaaccag ttccctgaaa ttattcccct
     4441 acttgactaa taagtatata aagacggtag gtattgattg taattctgta aatctatttc
     4501 ttaaacttct taaattctac ttttatagtt agtctttttt ttagttttaa aacaccagaa
     4561 cttagtttcg acggattcta gaactagtgg atcccccggg ctgcaggaat gtctgaattg
     4621 attaaagaaa acatgcacat gaaattgtac atggaaggca cagttgacaa ccaccacttc
     4681 aaatgcacat ctgaaggcga aggcaaacca tacgaaggca cacaaacaat gagaattaaa
     4741 gttgttgaag gcggcccatt gccattcgct ttcgacattt tggctacatc tttcttgtac
     4801 ggctctaaaa cattcattaa ccacacacaa ggcattccag acttcttcaa acaatctttc
     4861 ccagaaggct tcacatggga aagagttact acttacgaag acggcggcgt tttgactgct
     4921 actcaagaca cttctttgca agacggctgc ttgatttaca acgttaaaat tagaggcgtt
     4981 aacttcactt ctaacggccc agttatgcaa aaaaaaactt tgggctggga agctttcact
     5041 gaaactttgt acccagctga cggcggcttg gaaggcagaa acgacatggc tttgaaattg
     5101 gttggcggct ctcacttgat tgctaacatt aaaactactt acagatctaa aaaaccagct
     5161 aaaaacttga aaatgccagg cgtttactac gttgactaca gattggaaag aattaaagaa
     5221 gctaacaacg aaacttacgt tgaacaacac gaagttgctg ttgctagata ctgcgacttg
     5281 ccatctaaat tgggccacaa attgaacggc tccggtagtg gctccagatt cttggaagaa
     5341 ttgccagaag ttgctgaatc tttcaagaac ttcagagaag ctgttagatc tgaaggaaag
     5401 ttgacagaaa gagaaaagtt gttgatttct gttgcttgct ctgttgctgt tagatgcgac
     5461 gcttgcacaa gaagacacgc tgaagaagct ttggaagctg gaattactga aggtgaattg
     5521 gctgaagctg ctgctgttgc tgctttgatt agagctggtt ctgctatgaa cactgcttct
     5581 gctattttca gaggcggtgg tggcgagagt aaacggaata agccagggaa ggcgacaggt
     5641 aaaggtaaac cagttggtga taaatggctg gatgatgcag gtaaagattc aggagcgcca
     5701 attccagatc gcattgctga taagttgcgt gataaagaat ttaaaagctt cgacgatttt
     5761 cggaaggctg tatgggaaga ggtgtcgaaa gatcctgagc ttagtaaaaa tttaaaccca
     5821 agcaataagt ctagtgtttc aaaaggttat tctccgttta ctccaaagaa tcaacaggtc
     5881 ggagggagaa aagtctatga acttcatgct gacaagccaa ttagtcaagg tggtgaggtt
     5941 tatgacatgg ataatatccg agtgactaca cctaaacgac atatcgatat tcaccgaggt
     6001 aagtaaattc gatatcaagc ttatcgatac cgtcgacctc gagtcatgta attagttatg
     6061 tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga
     6121 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
     6181 tttatatttc aaatttttct tttttttctg tacagacacg tgtacgcatg taacattata
     6241 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccgagct
     6301 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcactcg
     6361 agggggggcc cggtaccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt
     6421 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6481 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6541 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6601 aatgaatcgg ccaacgcgcg gggagaggcg gtttgc
//
