LOCUS       sequence-374362-        8033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression SUMO7-SIM6 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8033)
  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 8033)
  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-SUMO7-6SIM
FEATURES             Location/Qualifiers
     source          1..8033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(106..125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      919..1507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1408..1427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1661..1678
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1795..1816
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1831..1836,1837..1854,1855..1861)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1869..1885
                     /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     1874..1896
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1893..1909
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1893..1909
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1930..1948
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1995..2661
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2531..2554
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             2713..2739
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(5739..5758)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5740..5758)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5765..5782)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5765..5781)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5774..5796)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5922..6377
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6009..6028)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6219..6240
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6508..7311)
                     /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(7312..7527)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7551..7570)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7670..7692
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7730..7748)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7816..7920
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7921..7989,7990..8033,1..748)
                     /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 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
       61 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      121 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      181 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
      241 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
      301 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
      361 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
      421 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
      481 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
      541 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
      601 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
      661 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
      721 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
      781 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
      841 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
      901 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      961 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1021 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1081 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1141 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1201 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1261 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1321 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1381 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1441 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1501 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1561 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1621 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     1681 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     1741 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     1801 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     1861 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     1921 caagctcgaa attaaccctc actaaaggga acaaaagctg gtaccgggcc ccccctcgag
     1981 tcagttcgag tttatcatta tcaatactgc catttcaaag aatacgtaaa taattaatag
     2041 tagtgatttt cctaacttta tttagtcaaa aaattagcct tttaattctg ctgtaacccg
     2101 tacatgccca aaataggggg cgggttacac agaatatata acatcgtagg tgtctgggtg
     2161 aacagtttat tcctggcatc cactaaatat aatggagccc gctttttaag ctggcatcca
     2221 gaaaaaaaaa gaatcccagc accaaaatat tgttttcttc accaaccatc agttcatagg
     2281 tccattctct tagcgcaact acagagaaca ggggcacaaa caggcaaaaa acgggcacaa
     2341 cctcaatgga gtgatgcaac ctgcctggag taaatgatga cacaaggcaa ttgacccacg
     2401 catgtatcta tctcattttc ttacaccttc tattaccttc tgctctctct gatttggaaa
     2461 aagctgaaaa aaaaggttga aaccagttcc ctgaaattat tcccctactt gactaataag
     2521 tatataaaga cggtaggtat tgattgtaat tctgtaaatc tatttcttaa acttcttaaa
     2581 ttctactttt atagttagtc ttttttttag ttttaaaaca ccaagaactt agtttcgaat
     2641 aaacacacat aaacaaacaa attaattaaa aaaatgtctt tgatcacaag tggagcgtca
     2701 ggggcacttc agttgccacc gttggaaaga ctaactctgg gctccgctgg atcagcttta
     2761 gccttgaaat tagcaggtct tgatatcctg atcactctgt acaagggtac cggttcatcc
     2821 ggatcttcag ggagttctgg ttcaagcatg tcagaagaga agcctaagga gggcgtgaaa
     2881 accgagaatg accacattaa cttgaaggtt gctggacagg atgggtccgt ggttcagttc
     2941 aaaattaaac gccacacacc gttgtcgaaa ctgatgaaag cgtacagcga acgccagggt
     3001 ctttctatgc gtcagattcg cttccgcttt gatggtcaac cgattaatga aacggacacg
     3061 ccggcacagc tggagatgga ggatgaagac actatcgacg tatttcagca gcagaccgtc
     3121 gtagggggct ccggcggaag cggggggtcg ggaggttcca tgagcgaaga gaaaccaaaa
     3181 gaaggagtta aaaccgaaaa cgaccatatc aacctgaagg tcgccggtca ggacggctca
     3241 gtcgtgcagt tcaagattaa acgtcatacc ccattaagta agctgatgaa ggcgtatagt
     3301 gagcgccaag gcttgtcgat gcgccagatt cgttttcgct ttgatggtca gccaatcaac
     3361 gaaaccgaca ccccggcgca gctggaaatg gaagatgaag ataccatcga tgtgttccaa
     3421 cagcaaacgg ttgttggcgg aagcggcggg agcggtggta gcgggggttc tatgtccgaa
     3481 gaaaaaccga aagagggtgt gaagactgaa aacgaccaca ttaatctgaa ggtggcgggc
     3541 caggacggtt ccgttgttca gtttaaaatc aaacgtcaca ccccgctgtc gaaactgatg
     3601 aaagcctata gcgaacgtca agggctgagc atgcgtcaaa ttcgttttcg ctttgatggt
     3661 cagccaatca acgaaaccga tacaccggcg cagctggaga tggaagacga ggacaccatt
     3721 gacgtcttcc agcaacaaac tgtacggtac aagggtaccg gttcatccgg atcttcaggg
     3781 agttctggtt caagcagatc tatgtcagaa gagaagccta aggaaggcgt gaaaaccgag
     3841 aatgaccaca ttaacttgaa ggttgctgga caggatgggt ccgtggttca gttcaagatt
     3901 aaacgccata caccgttgtc gaaactgatg aaggcgtaca gcgaacgtca gggtctttct
     3961 atgcgtcaga ttcgcttccg ctttgatggt caaccgatta atgaaacgga cacgccggca
     4021 cagctggaga tggaggatga ggacactatc gacgtatttc agcaacagac cgttgttgga
     4081 tcttcaggat caagcggttc atctggtagt tccatgtccg aagaaaaacc gaaggagggt
     4141 gtgaagactg aaaacgacca tattaatctg aaggtggcgg gccaggacgg ttccgttgtt
     4201 cagtttaaga tcaaacgcca caccccgctg tcgaaactga tgaaagccta tagtgaacgt
     4261 caagggctga gcatgcgtca aattcgtttt cgctttgatg gtcaaccaat caacgaaacc
     4321 gatacaccgg cgcagctgga aatggaagac gaggacacca ttgacgtctt ccagcaacaa
     4381 actgttgttg gttcctctgg aagctcagga tctagtgggt catcaactag tatgagtgaa
     4441 gaaaaaccca aggaaggcgt caagactgag aacgatcata ttaacctgaa agtcgcgggt
     4501 caagatggtt cagtggtgca gtttaaaatt aaacgccaca ccccgttgtc aaagctgatg
     4561 aaagcctata gtgaacgtca aggtctgtcg atgcgccaga tccgcttccg ctttgatggt
     4621 cagccaatca acgagacaga cactccggca caactggaga tggaagatga agacaccatc
     4681 gatgttttcc aacagcagac cgttgttggt tcatcaggtt catccggatc tagcgggtca
     4741 agtatgtctg aagaaaagcc gaaggaaggg gtaaagaccg aaaatgatca cattaatctg
     4801 aaagttgcgg gccaggatgg tagtgtagtg caatttaaaa ttaaacgtca caccccgttg
     4861 tctaaactga tgaaagcata tagcgaacgc caaggcctgt ctatgcgcca gattcgcttt
     4921 cgctttgacg ggcaaccaat caatgaaacg gacaccccgg cacaactgga aatggaagac
     4981 gaggacacta ttgacgtgtt tcagcaacag acaggttcat ccggatcttc agggagttct
     5041 ggttcaagca aggttgatgt aatagatttg accatagaat ctagttctga cgaggaagaa
     5101 gatcctcctg caaagagggg atcttcagga tcaagcggtt catctggtag ttcctctaga
     5161 aaggttgatg taatagattt gaccatagaa tctagttctg acgaggaaga agatcctcct
     5221 gcaaagaggg gtggttccgg gggttcaggg ggttctggtg gaagcggatc caaggttgat
     5281 gtaatagatt tgaccataga atctagttct gacgaggaag aagatcctcc tgcaaagagg
     5341 ggttcctctg gaagctcagg atctagtggg tcatcaaagg ttgatgtaat agatttgacc
     5401 atagaatcta gttctgacga ggaagaagat cctcctgcaa agaggggtgg ttccgggggt
     5461 tcagggggtt ctggtggaag caaggttgat gtaatagatt tgaccataga atctagttct
     5521 gacgaggaag aagatcctcc tgcaaagagg ggtggttccg ggggttcagg gggttctggt
     5581 ggaagcaagg ttgatgtaat agatttgacc atagaatcta gttctgacga ggaagaagat
     5641 cctcctgcaa agaggtaagg cgcgcctttt tttttatata tatatatata tatatataac
     5701 tgaataaaga gtatcatctt tcaaagagct ccaattcgcc ctatagtgag tcgtattaca
     5761 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     5821 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     5881 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     5941 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     6001 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     6061 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6121 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     6181 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     6241 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     6301 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     6361 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     6421 caccgcatag ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat
     6481 acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc
     6541 ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa
     6601 tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct
     6661 atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa
     6721 ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa
     6781 atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga
     6841 gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc
     6901 tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat
     6961 gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc
     7021 aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt
     7081 tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag
     7141 taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac
     7201 atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc
     7261 aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct
     7321 tcgtttcctg caggtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt
     7381 ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc
     7441 ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa
     7501 agaataaaaa aaaaatgatg aattgaaaag gtggtatggt gcactctcag tacaatctgc
     7561 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     7621 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     7681 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     7741 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     7801 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     7861 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     7921 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     7981 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gtt
//
