LOCUS Exported 7797 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression of human 6His-tagged SUMO3, SUMO-E1 and SUMO-E2. ACCESSION . VERSION . KEYWORDS pSUMO3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7797) AUTHORS Weber AR, Schuermann D, Schar P TITLE Versatile Recombinant SUMOylation System for the Production of SUMO-Modified Protein. JOURNAL PLoS One. 2014 Jul 9;9(7):e102157. doi: 10.1371/journal.pone.0102157. eCollection 2014. PUBMED 25007328 REFERENCE 2 (bases 1 to 7797) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7797 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /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)." CDS 100..117 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 139..414 /codon_start=1 /product="human small ubiquitin-related modifier 3" /label=SUMO3 /translation="MSEEKPKEGVKTENDHINLKVAGQDGSVVQFKIKRHTPLSKLMKA YCERQGLSMRQIRFRFDGQPINETDTPAQLEMEDEDTIDVFQQQTGG" promoter 560..578 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 579..603 /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)." CDS 1135..1179 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" terminator 1231..1278 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter 1363..1381 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1382..1406 /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)." terminator 4495..4542 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(4713..5504) /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" promoter complement(5505..5596) /gene="bla" /label=AmpR promoter rep_origin complement(5644..6382) /direction=LEFT /label=CloDF13 ori /note="Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins." CDS complement(6592..7674) /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 7636..7655 /label=LacI-R /note="LacI, reverse primer" promoter complement(7675..7752) /gene="lacI" /label=lacI promoter ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt 61 tgtttaactt taataaggag atataccatg ggcagcagcc atcaccatca tcaccacagc 121 caggatctgg ttccgcgtat gtccgaggag aagcccaagg agggtgtgaa gacagagaat 181 gaccacatca acctgaaggt ggcagggcag gacggctccg tggtgcagtt caagatcaag 241 aggcacacgc cgctgagcaa gctgatgaag gcctactgcg agaggcaggg cttgtcaatg 301 aggcagatca gattcaggtt cgacgggcag ccaatcaatg aaactgacac tccagcacag 361 ctggagatgg aggacgagga caccatcgac gtgttccagc agcagacggg aggttagctc 421 gagcggccgc atcgtgactg actgacgctg caggtcgaca agcttgcggc cgcataatgc 481 ttaacccctt ggggcctcta aacgggtctt gaggggtttt ttgcatgcat cgtattgtac 541 acggccgcat aatcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat 601 tccccatctt agtatattag ttaagtataa gaaggagata tacatatgtc ggggatcgcc 661 ctcagcagac tcgcccagga gaggaaagca tggaggaaag accacccatt tggtttcgtg 721 gctgtcccaa caaaaaatcc cgatggcacg atgaacctca tgaactggga gtgcgccatt 781 ccgggaaaga aagggactcc gtgggaagga ggcttgttta aactacggat gcttttcaaa 841 gatgattatc catcttcgcc accaaaatgt aaattcgaac caccattatt tcacccgaat 901 gtgtaccctt cggggacagt gtgcctgtcc atcttagagg aggacaagga ctggaggcca 961 gccatcacaa tcaaacagat cctattagga atacaggaac ttctaaatga accaaatatc 1021 caagacccag ctcaagcaga ggcctacacg atttactgcc aaaacagagt ggagtacgag 1081 aaaagggtcc gagcacaagc caagaagttt gcgccctcat aactcgagtc tggtaaagaa 1141 accgctgctg cgaaatttga acgccagcac atggactcgt ctactagcgc agcttaatta 1201 acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg 1261 gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacggtac cattaagtcg 1321 aacagaaagt aatcgtattg tacacggccg cataatcgaa attaatacga ctcactatag 1381 gggaattgtg agcggataac aattccccat cttagtatat tagttaagta taagaaggag 1441 atatacatat ggcagatctc aattcccggg tcgacatggc actgtcgcgg gggctgcccc 1501 gggagctggc tgaggcggtg gccgggggcc gggtgctggt ggtgggggcg ggcggcatcg 1561 gctgcgagct cctcaagaat ctcgtgctca ccggtttctc ccacatcgac ctgattgatc 1621 tggatactat tgatgtaagc aacctcaaca gacagttttt gtttcaaaag aaacatgttg 1681 gaagatcaaa ggcacaggtt gccaaggaaa gtgtactgca gttttacccg aaagctaata 1741 tcgttgccta ccatgacagc atcatgaacc ctgactataa tgtggaattt ttccgacagt 1801 ttatactggt tatgaatgct ttagataaca gagctgcccg aaaccatgtt aatagaatgt 1861 gcctggcagc tgatgttcct cttattgaaa gtggaacagc tgggtatctt ggacaagtaa 1921 ctactatcaa aaagggtgtg accgagtgtt atgagtgtca tcctaagccg acccagagaa 1981 cctttcctgg ctgtacaatt cgtaacacac cttcagaacc tatacattgc atcgtttggg 2041 caaagtactt gttcaaccag ttgtttgggg aagaagatgc tgatcaagaa gtatctcctg 2101 acagagctga ccctgaagct gcctgggaac caacggaagc cgaagccaga gctagagcat 2161 ctaatgaaga tggtgacatt aaacgtattt ctactaagga atgggctaaa tcaactggat 2221 atgatccagt taaacttttt accaagcttt ttaaagatga catcaggtat ctgttgacaa 2281 tggacaaact atggcggaaa aggaaacctc cagttccgtt ggactgggct gaagtacaaa 2341 gtcaaggaga agaaacgaat gcatcagatc aacagaatga accccagtta ggcctgaaag 2401 accagcaggt tctagatgta aagagctatg cacgtctttt ttcaaagagc atcgagactt 2461 tgagagttca tttagcagaa aagggggatg gagctgagct catatgggat aaggatgacc 2521 catctgcaat ggattttgtc acctctgctg caaacctcag gatgcatatt ttcagtatga 2581 atatgaagag tagatttgat atcaaatcaa tggcagggaa cattattcct gctattgcta 2641 ctactaatgc agtaattgct gggttgatag tattggaagg attgaagatt ttatcaggaa 2701 aaatagacca gtgcagaaca atttttttga ataaacaacc aaacccaaga aagaagcttc 2761 ttgtgccttg tgcactggat cctcccaacc ccaattgtta tgtatgtgcc agcaagccag 2821 aggtgactgt gcggctgaat gtccataaag tgactgttct caccttacaa gacaagatag 2881 tgaaagaaaa atttgctatg gtagcaccag atgtccaaat tgaagatggg aaaggaacaa 2941 tcctaatatc ttccgaagag ggagagacgg aagctaataa tcacaagaag ttgtcagaat 3001 ttggaattag aaatggcagc cggcttcaag cagatgactt cctccaggac tatactttat 3061 tgatcaacat ccttcatagt gaagacctag gaaaggacgt tgaatttgaa gttgttggtg 3121 atgctccgga aaaagtgggg cccaaacaag ctgaagatgc tgccaaaagc ataaccaatg 3181 gcagtgatga tggagctcag ccctccacct ccacagctca agagcaagat gacgttctca 3241 tagttgattc ggatgaagaa gattcttcaa ataatgccga cgtcagtgaa gaagagagaa 3301 gccgcaagag gaaattagat gagaaagaga atctcagtgc aaagaggtca cgtatagaac 3361 agaaggaaga gcttgatgat gtcatagcat tagattgagc ggccgcatta aaaggaggaa 3421 ttaactatgg tggagaagga ggaggctggc ggcggcatta gcgaggagga ggcggcacag 3481 tatgaccggc agatccgcct gtggggactg gaggcccaga aacggctgcg ggcctctcgg 3541 gtgcttcttg tcggcttgaa aggacttggg gctgaaattg ccaagaatct catcttggca 3601 ggagtgaaag gactgaccat gctggatcac gaacaggtaa ctccagaaga tcccggagct 3661 cagttcttga ttcgtactgg gtctgttggc cgaaataggg ctgaagcctc tttggagcga 3721 gctcagaatc tcaaccccat ggtggatgtg aaggtggaca ctgaggatat agagaagaaa 3781 ccagagtcat ttttcactca attcgatgct gtgtgtctga cttgctgctc cagggatgtc 3841 atagttaaag ttgaccagat ctgtcacaaa aatagcatca agttctttac aggagatgtt 3901 tttggctacc atggatacac atttgccaat ctaggagagc atgagtttgt agaggagaaa 3961 actaaagttg ccaaagttag ccaaggagta gaagatgggc ccgacaccaa gagagcaaaa 4021 cttgattctt ctgagacaac gatggtcaaa aagaaggtgg tcttctgccc tgttaaagaa 4081 gccctggagg tggactggag cagtgagaaa gcaaaggctg ctctgaagcg cacgacctcc 4141 gactactttc tccttcaagt gctcttaaag ttccgtacag ataaaggaag agatcccagt 4201 tctgatacat atgaggaaga ttctgagttg ttgctccaga tacgaaatga tgtgcttgac 4261 tcactgggta ttagtcctga cctgcttcct gaggactttg tcaggtactg cttctccgag 4321 atggccccag tgtgtgcggt ggttggaggg attttggcac aggaaattgt gaaggccctg 4381 tctcagcggg accctcctca caacaacttc ttcttcttcg atggcatgaa ggggaatggg 4441 attgtggagt gccttggccc caagggctag gctgctgcca ccgctgagca ataactagca 4501 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt 4561 gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca 4621 taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcatcgtg gccggatctt 4681 gcggcccctc ggcttgaacg aattgttaga cattatttgc cgactacctt ggtgatctcg 4741 cctttcacgt agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct 4801 tcttgtccaa gataagcctg tctagcttca agtatgacgg gctgatactg ggccggcagg 4861 cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt tactgcgctg 4921 taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca gtcgggcggc 4981 gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc aggaaccgga 5041 tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct tgcttttgtc 5101 agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc aagaatgtca 5161 ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca cggaatgatg 5221 tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa 5281 gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg 5341 gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca ggccgccatc cactgcggag 5401 ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc 5461 tctgatagtt gagtcgatac ttcggcgatc accgcttccc tcatactctt cctttttcaa 5521 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 5581 tagaaaaata aacaaatagc tagctcactc ggtcgctacg ctccgggcgt gagactgcgg 5641 cgggcgctgc ggacacatac aaagttaccc acagattccg tggataagca ggggactaac 5701 atgtgaggca aaacagcagg gccgcgccgg tggcgttttt ccataggctc cgccctcctg 5761 ccagagttca cataaacaga cgcttttccg gtgcatctgt gggagccgtg aggctcaacc 5821 atgaatctga cagtacgggc gaaacccgac aggacttaaa gatccccacc gtttccggcg 5881 ggtcgctccc tcttgcgctc tcctgttccg accctgccgt ttaccggata cctgttccgc 5941 ctttctccct tacgggaagt gtggcgcttt ctcatagctc acacactggt atctcggctc 6001 ggtgtaggtc gttcgctcca agctgggctg taagcaagaa ctccccgttc agcccgactg 6061 ctgcgcctta tccggtaact gttcacttga gtccaacccg gaaaagcacg gtaaaacgcc 6121 actggcagca gccattggta actgggagtt cgcagaggat ttgtttagct aaacacgcgg 6181 ttgctcttga agtgtgcgcc aaagtccggc tacactggaa ggacagattt ggttgctgtg 6241 ctctgcgaaa gccagttacc acggttaagc agttccccaa ctgacttaac cttcgatcaa 6301 accacctccc caggtggttt tttcgtttac agggcaaaag attacgcgca gaaaaaaagg 6361 atctcaagaa gatcctttga tcttttctac tgaaccgctc tagatttcag tgcaatttat 6421 ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgtta 6481 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt 6541 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc 6601 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 6661 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa 6721 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt 6781 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct 6841 gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc 6901 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg 6961 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc 7021 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc 7081 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg 7141 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat 7201 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc 7261 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac 7321 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac 7381 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat 7441 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt 7501 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc 7561 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt 7621 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac 7681 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca 7741 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaaat //