LOCUS       sequence-374354-        6379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression dimer-1SUMO component in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6379)
  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 6379)
  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: pRS306-PTDH3-Dimer-1SUMO
FEATURES             Location/Qualifiers
     source          1..6379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(177..196)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      990..1578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1479..1498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1732..1749
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1866..1887
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1902..1907,1908..1925,1926..1932)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1940..1956
                     /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     1945..1967
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1964..1980
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1964..1980
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2001..2019
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2074..2717
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2611..2634
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     2722..2738
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2722..2738
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      3886..4133
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3902..3920)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4156..4175)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4157..4175)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4182..4199)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4182..4198)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4191..4213)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4339..4794
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4426..4445)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4636..4657
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4925..5728)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(5729..5944)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5968..5987)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6087..6109
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6147..6165)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6233..6337
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6338..6379,1..27,28..819)
                     /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"
ORIGIN
        1 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
       61 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
      121 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
      181 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
      241 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
      301 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
      361 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
      421 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
      481 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
      541 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
      601 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
      661 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
      721 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
      781 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
      841 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
      901 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
      961 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1021 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1081 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1141 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1201 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1261 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     1321 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     1381 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     1441 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     1501 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     1561 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     1621 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     1681 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     1741 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     1801 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     1861 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     1921 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     1981 catgattacg ccaagctcga aattaaccct cactaaaggg aacaaaagct ggtaccgggc
     2041 cccccctcga gacaaaagct ggagctcagt ttatcattat caatactcgc catttcaaag
     2101 aatacgtaaa taattaatag tagtgatttt cctaacttta tttagtcaaa aaattagcct
     2161 tttaattctg ctgtaacccg tacatgccca aaataggggg cgggttacac agaatatata
     2221 acatcgtagg tgtctgggtg aacagtttat tcctggcatc cactaaatat aatggagccc
     2281 gctttttaag ctggcatcca gaaaaaaaaa gaatcccagc accaaaatat tgttttcttc
     2341 accaaccatc agttcatagg tccattctct tagcgcaact acagagaaca ggggcacaaa
     2401 caggcaaaaa acgggcacaa cctcaatgga gtgatgcaac ctgcctggag taaatgatga
     2461 cacaaggcaa ttgacccacg catgtatcta tctcattttc ttacaccttc tattaccttc
     2521 tgctctctct gatttggaaa aagctgaaaa aaaaggttga aaccagttcc ctgaaattat
     2581 tcccctactt gactaataag tatataaaga cggtaggtat tgattgtaat tctgtaaatc
     2641 tatttcttaa acttcttaaa ttctactttt atagttagtc ttttttttag ttttaaaaca
     2701 ccagaactta gtttcgacgg atctagaact agtggatccc ccgggctgca ggacaaaatg
     2761 ttggctgaaa agttggctgg attggacatt aactctggcg aactgaaaca tagtattagt
     2821 gattataccg aggctgaatt tctggagttt gtaaaaaaaa tatgtagagc tgaaggtgct
     2881 actgaagagg atgacaataa attagtgaga gagtttgctc gattaactga gcacccagac
     2941 ggttcagatc tgatttatta tcctcgcgat gacagggaag atagtcctga agggattgtc
     3001 aaggaaatta aagaatggcg agctgctaac ggtaagtcag gatttaaaca gggcggcgga
     3061 cgtacgggtg gcatgtcaga agagaagcct aaggagggcg tgaaaaccga gaatgaccac
     3121 attaacttga aggttgctgg acaggatggg tccgtggttc agttcaaaat taaacgccac
     3181 acaccgttgt cgaaactgat gaaagcgtac agcgaacgcc agggtctttc tatgcgtcag
     3241 attcgcttcc gctttgatgg tcaaccgatt aatgaaacgg acacgccggc acagctggag
     3301 atggaggatg aagacactat cgacgtattt cagcagcaga cctctagtgc tagcagttct
     3361 ttgtctcaag cttctgctga cttggaagct acattgagac acaaattgac agttatggat
     3421 tctcaaatta acggagcttc tagagctttg gacgacgtta gaaacagaca acaagacgtt
     3481 agaatgacag ctaacagaaa agttgaacaa ttgcaacaag aatacactga aatgaaagct
     3541 ttgttggacg cttctgaaac tacttctact agaaagatta aagaagaaga aaagagagtt
     3601 aactctaaat tcgacactat ttaccaagat ttgttgaaga agaagtctga aattcaaact
     3661 ttgaaagaag aaattgaaca atctttgact aaaagagacg aattcgaatt cttggaaaag
     3721 gcttctaaat tgagaggaat ttctactaaa ccagtttaca ttccagaagt tgaattgaac
     3781 cacaaattga ttaaaggaat tcaccaatct actattgact tgaagaacga attgaaacaa
     3841 tgcattggta gattgcaaga attgactcca taagtcgacc tcgagtcatg taattagtta
     3901 tgtcacgctt acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta
     3961 gacaacctga agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt
     4021 tatttatatt tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta
     4081 tactgaaaac cttgcttgag aaggttttgg gacgctcgaa ggctttaatt tgcggccggt
     4141 acgagctcca attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa
     4201 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     4261 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4321 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4381 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4441 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4501 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4561 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4621 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4681 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4741 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
     4801 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagggt aataactgat
     4861 ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag
     4921 ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac
     4981 gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa
     5041 tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc
     5101 ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc
     5161 cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt
     5221 caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta
     5281 acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa
     5341 caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt
     5401 atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt
     5461 cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca
     5521 tggaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg
     5581 cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg
     5641 tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag
     5701 cacgttcctt atatgtagct ttcgacatga tttatcttcg tttcctgcag gtttttgttc
     5761 tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta
     5821 tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg
     5881 aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat
     5941 tgaaaaggtg gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6001 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     6061 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     6121 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     6181 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6241 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6301 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6361 cgcccttatt ccctttttt
//
