LOCUS       sequence-459202-        5689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of E67R SUMO1 protein under T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5689)
  AUTHORS   Kim J, Qin S, Zhou HX, Rosen MK
  TITLE     Surface Charge Can Modulate Phase Separation of Multidomain 
            Proteins.
  JOURNAL   J Am Chem Soc. 2024 Feb 7;146(5):3383-3395. doi: 
            10.1021/jacs.3c12789. Epub 2024 Jan 23.
  PUBMED    38262618
REFERENCE   2  (bases 1 to 5689)
  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: 
            "10.1021/jacs.3c12789"; journalName: "J Am Chem Soc"; date: 
            "2024-02-7- 7"; volume: "146"; issue: "5"; pages: "3383-3395"
COMMENT     Sequence Label: E67R SUMO1
FEATURES             Location/Qualifiers
     source          1..5689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4..22)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        90..194
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(195..263,264..1055)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(413..432)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1226..1814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1715..1734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1968..1985
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2000..2139
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2065..2084)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2225..2247
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2241..2432)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3744..4826)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4788..4807
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4827..4904)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5110..5129
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5213..5232
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5213..5231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5232..5256
                     /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)."
     RBS             5271..5293
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5307..5330
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5331..5351
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
       61 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
      121 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      181 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
      241 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
      301 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
      361 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
      421 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
      481 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
      541 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
      601 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
      661 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
      721 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
      781 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
      841 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
      901 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
      961 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1021 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1081 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1141 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1201 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1261 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1321 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     1381 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1441 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1501 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1561 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1621 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1681 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1741 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1801 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     1861 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     1921 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     1981 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2041 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     2101 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     2161 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2221 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     2281 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     2341 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     2401 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     2461 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     2521 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     2581 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     2641 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     2701 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     2761 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     2821 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     2881 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg
     2941 ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt
     3001 ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc
     3061 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     3121 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
     3181 cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg
     3241 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
     3301 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     3361 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     3421 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     3481 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     3541 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     3601 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     3661 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     3721 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3781 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     3841 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     3901 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     3961 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     4021 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     4081 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     4141 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     4201 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     4261 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     4321 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     4381 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     4441 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     4501 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     4561 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     4621 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     4681 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     4741 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     4801 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     4861 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     4921 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     4981 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     5041 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     5101 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     5161 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
     5221 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
     5281 tttaagaagg agatatacat atcatgcacc accatcacca ccatcatcac gaaaacctgt
     5341 attttcaggg catgcatgaa ttcggcggct catggggtgg ttctatgagt gatcaggaag
     5401 ccaaaccctc aacggaggat cttggtgata aaaaagaagg ggagtatatt aaactgaaag
     5461 tgattggaca ggacagtagc gagattcatt tcaaagttaa gatgactacg catctgaaaa
     5521 agctgaagga atcgtattgc cagcgccagg gtgttccgat gaacagctta cgcttcctgt
     5581 tccgtggcca gcgcatcgcg gacaaccata ctcctaaaga gctgggtatg gaagaggagg
     5641 acgtgattga agtataccag gaacagaccg gcggataaca aagcccgaa
//
