LOCUS       sequence-374360-        8957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression SUMO10-SIM6 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8957)
  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 8957)
  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-SUMO10-6SIM
FEATURES             Location/Qualifiers
     source          1..8957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(107..126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      920..1508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1409..1428
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1662..1679
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1796..1817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1832..1837,1838..1855,1856..1862)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1870..1886
                     /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     1875..1897
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1894..1910
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1894..1910
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1931..1949
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1996..2662
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2532..2555
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             2714..2740
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(6664..6683)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6665..6683)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6690..6707)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6690..6706)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6699..6721)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6847..7302
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6934..6953)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7144..7165
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7433..8236)
                     /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(8237..8452)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(8476..8495)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8595..8617
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8655..8673)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8741..8845
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8846..8914,8915..8957,1..749)
                     /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 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
       61 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      121 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      181 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      241 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      301 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      361 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      421 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      481 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      541 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      601 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      661 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      721 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
      781 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
      841 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
      901 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
      961 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1021 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1081 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1141 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1201 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1261 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1321 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1381 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1441 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1501 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1561 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1621 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1681 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1741 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     1801 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     1861 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     1921 ccaagctcga aattaaccct cactaaaggg aacaaaagct ggtaccgggc cccccctcga
     1981 gtcagttcga gtttatcatt atcaatactg ccatttcaaa gaatacgtaa ataattaata
     2041 gtagtgattt tcctaacttt atttagtcaa aaaattagcc ttttaattct gctgtaaccc
     2101 gtacatgccc aaaatagggg gcgggttaca cagaatatat aacatcgtag gtgtctgggt
     2161 gaacagttta ttcctggcat ccactaaata taatggagcc cgctttttaa gctggcatcc
     2221 agaaaaaaaa agaatcccag caccaaaata ttgttttctt caccaaccat cagttcatag
     2281 gtccattctc ttagcgcaac tacagagaac aggggcacaa acaggcaaaa aacgggcaca
     2341 acctcaatgg agtgatgcaa cctgcctgga gtaaatgatg acacaaggca attgacccac
     2401 gcatgtatct atctcatttt cttacacctt ctattacctt ctgctctctc tgatttggaa
     2461 aaagctgaaa aaaaaggttg aaaccagttc cctgaaatta ttcccctact tgactaataa
     2521 gtatataaag acggtaggta ttgattgtaa ttctgtaaat ctatttctta aacttcttaa
     2581 attctacttt tatagttagt ctttttttta gttttaaaac accaagaact tagtttcgaa
     2641 taaacacaca taaacaaaca aattaattaa aaaaatgtct ttgatcacaa gtggagcgtc
     2701 aggggcactt cagttgccac cgttggaaag actaactctg ggctccgctg gatcagcttt
     2761 agccttgaaa ttagcaggtc ttgatatcct gatcactctg tacaagggta ccggttcatc
     2821 cggatcttca gggagttctg gttcaagcat gtcagaagag aagcctaagg agggcgtgaa
     2881 aaccgagaat gaccacatta acttgaaggt tgctggacag gatgggtccg tggttcagtt
     2941 caaaattaaa cgccacacac cgttgtcgaa actgatgaaa gcgtacagcg aacgccaggg
     3001 tctttctatg cgtcagattc gcttccgctt tgatggtcaa ccgattaatg aaacggacac
     3061 gccggcacag ctggagatgg aggatgaaga cactatcgac gtatttcagc agcagaccgt
     3121 cgtagggggc tccggcggaa gcggggggtc gggaggttcc atgagcgaag agaaaccaaa
     3181 agaaggagtt aaaaccgaaa acgaccatat caacctgaag gtcgccggtc aggacggctc
     3241 agtcgtgcag ttcaagatta aacgtcatac cccattaagt aagctgatga aggcgtatag
     3301 tgagcgccaa ggcttgtcga tgcgccagat tcgttttcgc tttgatggtc agccaatcaa
     3361 cgaaaccgac accccggcgc agctggaaat ggaagatgaa gataccatcg atgtgttcca
     3421 acagcaaacg gttgttggcg gaagcggcgg gagcggtggt agcgggggtt ctatgtccga
     3481 agaaaaaccg aaagagggtg tgaagactga aaacgaccac attaatctga aggtggcggg
     3541 ccaggacggt tccgttgttc agtttaaaat caaacgtcac accccgctgt cgaaactgat
     3601 gaaagcctat agcgaacgtc aagggctgag catgcgtcaa attcgttttc gctttgatgg
     3661 tcagccaatc aacgaaaccg atacaccggc gcagctggag atggaagacg aggacaccat
     3721 tgacgtcttc cagcaacaaa ctgtagtggg tggctccggt ggctctggtg ggtctggtgg
     3781 ctcgatgagt gaggaaaaac ccaaagaagg cgtcaagact gagaacgatc atattaacct
     3841 gaaagtcgcg ggtcaagatg gttcagtggt gcagttcaaa attaaacgcc acaccccgct
     3901 gtcaaagctg atgaaagcct atagtgagcg tcaaggtctg tcgatgcgcc agatccgctt
     3961 tcgctttgat ggtcagccaa tcaacgagac agacactccg gcacaactgg agatggaaga
     4021 tgaagacacc atcgatgttt ttcaacagca gaccgtggtg ggagggagcg gcggtagcgg
     4081 gggatcaggt ggtagtatgt ctgaggaaaa gccgaaggaa ggggtaaaga ccgaaaatga
     4141 tcacattaat ctgaaagttg cgggccaaga tggtagtgta gtgcaattta aaatcaagcg
     4201 tcacaccccg ttatctaaac tgatgaaagc atatagcgaa cgccaaggcc tgtctatgcg
     4261 ccagattcgc tttcgctttg acgggcaacc aattaatgaa acggacaccc cggcacaact
     4321 ggaaatggaa gacgaggata ctattgacgt gtttcagcaa cagacagtcg taggttcatc
     4381 cggatcttca gggagttctg gttcaagcat gagcgaagag aaacctaaag aaggagttaa
     4441 aaccgaaaat gaccatatca acctgaaggt ggccggccag gacggctcag tcgtgcagtt
     4501 caagatcaaa cgtcataccc cattaagtaa gctgatgaag gcgtatagtg agcgccaagg
     4561 cttgtcgatg cgccagattc gtttccgctt tgacggtcag ccaatcaacg aaaccgacac
     4621 cccggcgcag ctggaaatgg aagacgaaga tactatcgat gtgttccaac agcaaacggt
     4681 cgtaggatct tcaggatcaa gcggttcatc tggtagttcc agatctatgt cagaagagaa
     4741 gcctaaggaa ggcgtgaaaa ccgagaatga ccacattaac ttgaaggttg ctggacagga
     4801 tgggtccgtg gttcagttca agattaaacg ccatacaccg ttgtcgaaac tgatgaaggc
     4861 gtacagcgaa cgtcagggtc tttctatgcg tcagattcgc ttccgctttg atggtcaacc
     4921 gattaatgaa acggacacgc cggcacagct ggagatggag gatgaggaca ctatcgacgt
     4981 atttcagcaa cagaccgttg ttggatcttc aggatcaagc ggttcatctg gtagttccat
     5041 gtccgaagaa aaaccgaagg agggtgtgaa gactgaaaac gaccatatta atctgaaggt
     5101 ggcgggccag gacggttccg ttgttcagtt taagatcaaa cgccacaccc cgctgtcgaa
     5161 actgatgaaa gcctatagtg aacgtcaagg gctgagcatg cgtcaaattc gttttcgctt
     5221 tgatggtcaa ccaatcaacg aaaccgatac accggcgcag ctggaaatgg aagacgagga
     5281 caccattgac gtcttccagc aacaaactgt tgttggttcc tctggaagct caggatctag
     5341 tgggtcatca actagtatga gtgaagaaaa acccaaggaa ggcgtcaaga ctgagaacga
     5401 tcatattaac ctgaaagtcg cgggtcaaga tggttcagtg gtgcagttta aaattaaacg
     5461 ccacaccccg ttgtcaaagc tgatgaaagc ctatagtgaa cgtcaaggtc tgtcgatgcg
     5521 ccagatccgc ttccgctttg atggtcagcc aatcaacgag acagacactc cggcacaact
     5581 ggagatggaa gatgaagaca ccatcgatgt tttccaacag cagaccgttg ttggttcatc
     5641 aggttcatcc ggatctagcg ggtcaagtat gtctgaagaa aagccgaagg aaggggtaaa
     5701 gaccgaaaat gatcacatta atctgaaagt tgcgggccag gatggtagtg tagtgcaatt
     5761 taaaattaaa cgtcacaccc cgttgtctaa actgatgaaa gcatatagcg aacgccaagg
     5821 cctgtctatg cgccagattc gctttcgctt tgacgggcaa ccaatcaatg aaacggacac
     5881 cccggcacaa ctggaaatgg aagacgagga cactattgac gtgtttcagc aacagacagg
     5941 ttcatccgga tcttcaggga gttctggttc aagcaaggtt gatgtaatag atttgaccat
     6001 agaatctagt tctgacgagg aagaagatcc tcctgcaaag aggggatctt caggatcaag
     6061 cggttcatct ggtagttcct ctagaaaggt tgatgtaata gatttgacca tagaatctag
     6121 ttctgacgag gaagaagatc ctcctgcaaa gaggggtggt tccgggggtt cagggggttc
     6181 tggtggaagc ggatccaagg ttgatgtaat agatttgacc atagaatcta gttctgacga
     6241 ggaagaagat cctcctgcaa agaggggttc ctctggaagc tcaggatcta gtgggtcatc
     6301 aaaggttgat gtaatagatt tgaccataga atctagttct gacgaggaag aagatcctcc
     6361 tgcaaagagg ggtggttccg ggggttcagg gggttctggt ggaagcaagg ttgatgtaat
     6421 agatttgacc atagaatcta gttctgacga ggaagaagat cctcctgcaa agaggggtgg
     6481 ttccgggggt tcagggggtt ctggtggaag caaggttgat gtaatagatt tgaccataga
     6541 atctagttct gacgaggaag aagatcctcc tgcaaagagg taaggcgcgc cttttttttt
     6601 atatatatat atatatatat ataactgaat aaagagtatc atctttcaaa gagctccaat
     6661 tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg
     6721 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     6781 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     6841 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     6901 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     6961 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     7021 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     7081 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     7141 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     7201 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     7261 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     7321 ccttacgcat ctgtgcggta tttcacaccg catagggtaa taactgatat aattaaattg
     7381 aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt
     7441 gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta
     7501 ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg
     7561 tagagaccac atcatccacg gttctatact gttgacccaa tgcgtctccc ttgtcatcta
     7621 aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc
     7681 tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct
     7741 tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc
     7801 ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc
     7861 ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag
     7921 agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa
     7981 aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag
     8041 tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact
     8101 ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt
     8161 gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat
     8221 atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg
     8281 ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc
     8341 taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat
     8401 ttcaaggaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aaaaggtggt
     8461 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     8521 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     8581 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     8641 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     8701 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     8761 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     8821 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     8881 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     8941 agatgctgaa gatcagt
//
