LOCUS Exported 7815 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression of human 6His-tagged SUMO1, SUMO-E1 and SUMO-E2. ACCESSION . VERSION . KEYWORDS pSUMO1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7815) 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 7815) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7815 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 2..20 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 21..45 /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 101..118 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" promoter 579..597 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 598..622 /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 1154..1198 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" terminator 1250..1297 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter 1382..1400 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1401..1425 /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 4514..4561 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(4732..5523) /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(5524..5615) /gene="bla" /label=AmpR promoter rep_origin complement(5663..6401) /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(6611..7693) /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 7655..7674 /label=LacI-R /note="LacI, reverse primer" promoter complement(7694..7771) /gene="lacI" /label=lacI promoter ORIGIN 1 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctgt agaaataatt 61 ttgtttaact ttaataagga gatataccat gggcagcagc catcaccatc atcaccacag 121 ccaggatctg gttccgcgta tgtctgacca ggaggccaaa ccttcaactg aggacttggg 181 ggataagaag gaaggtgaat atattaaact caaagtcatt ggacaggata gcagtgagat 241 tcacttcaaa gtgaaaatga caacacatct caagaaactc aaagaatcat actgtcaaag 301 acagggtgtt ccaatgaatt cactcaggtt tctctttgag ggtcagagaa ttgctgataa 361 tcatactcca aaagaactgg gaatggagga agaagatgtg attgaagttt atcaggaaca 421 aacggggggt tagggatccc cgggaattca tcgtgactga ctgacgctgc aggtcgacaa 481 gcttgcggcc gcataatgct taaccccttg gggcctctaa acgggtcttg aggggttttt 541 tgcatgcatc gtattgtaca cggccgcata atcgaaatta atacgactca ctatagggga 601 attgtgagcg gataacaatt ccccatctta gtatattagt taagtataag aaggagatat 661 acatatgtcg gggatcgccc tcagcagact cgcccaggag aggaaagcat ggaggaaaga 721 ccacccattt ggtttcgtgg ctgtcccaac aaaaaatccc gatggcacga tgaacctcat 781 gaactgggag tgcgccattc cgggaaagaa agggactccg tgggaaggag gcttgtttaa 841 actacggatg cttttcaaag atgattatcc atcttcgcca ccaaaatgta aattcgaacc 901 accattattt cacccgaatg tgtacccttc ggggacagtg tgcctgtcca tcttagagga 961 ggacaaggac tggaggccag ccatcacaat caaacagatc ctattaggaa tacaggaact 1021 tctaaatgaa ccaaatatcc aagacccagc tcaagcagag gcctacacga tttactgcca 1081 aaacagagtg gagtacgaga aaagggtccg agcacaagcc aagaagtttg cgccctcata 1141 actcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc 1201 tactagcgca gcttaattaa cctaggctgc tgccaccgct gagcaataac tagcataacc 1261 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg catttgagaa 1321 gcacggtacc attaagtcga acagaaagta atcgtattgt acacggccgc ataatcgaaa 1381 ttaatacgac tcactatagg ggaattgtga gcggataaca attccccatc ttagtatatt 1441 agttaagtat aagaaggaga tatacatatg gcagatctca attcccgggt cgacatggca 1501 ctgtcgcggg ggctgccccg ggagctggct gaggcggtgg ccgggggccg ggtgctggtg 1561 gtgggggcgg gcggcatcgg ctgcgagctc ctcaagaatc tcgtgctcac cggtttctcc 1621 cacatcgacc tgattgatct ggatactatt gatgtaagca acctcaacag acagtttttg 1681 tttcaaaaga aacatgttgg aagatcaaag gcacaggttg ccaaggaaag tgtactgcag 1741 ttttacccga aagctaatat cgttgcctac catgacagca tcatgaaccc tgactataat 1801 gtggaatttt tccgacagtt tatactggtt atgaatgctt tagataacag agctgcccga 1861 aaccatgtta atagaatgtg cctggcagct gatgttcctc ttattgaaag tggaacagct 1921 gggtatcttg gacaagtaac tactatcaaa aagggtgtga ccgagtgtta tgagtgtcat 1981 cctaagccga cccagagaac ctttcctggc tgtacaattc gtaacacacc ttcagaacct 2041 atacattgca tcgtttgggc aaagtacttg ttcaaccagt tgtttgggga agaagatgct 2101 gatcaagaag tatctcctga cagagctgac cctgaagctg cctgggaacc aacggaagcc 2161 gaagccagag ctagagcatc taatgaagat ggtgacatta aacgtatttc tactaaggaa 2221 tgggctaaat caactggata tgatccagtt aaacttttta ccaagctttt taaagatgac 2281 atcaggtatc tgttgacaat ggacaaacta tggcggaaaa ggaaacctcc agttccgttg 2341 gactgggctg aagtacaaag tcaaggagaa gaaacgaatg catcagatca acagaatgaa 2401 ccccagttag gcctgaaaga ccagcaggtt ctagatgtaa agagctatgc acgtcttttt 2461 tcaaagagca tcgagacttt gagagttcat ttagcagaaa agggggatgg agctgagctc 2521 atatgggata aggatgaccc atctgcaatg gattttgtca cctctgctgc aaacctcagg 2581 atgcatattt tcagtatgaa tatgaagagt agatttgata tcaaatcaat ggcagggaac 2641 attattcctg ctattgctac tactaatgca gtaattgctg ggttgatagt attggaagga 2701 ttgaagattt tatcaggaaa aatagaccag tgcagaacaa tttttttgaa taaacaacca 2761 aacccaagaa agaagcttct tgtgccttgt gcactggatc ctcccaaccc caattgttat 2821 gtatgtgcca gcaagccaga ggtgactgtg cggctgaatg tccataaagt gactgttctc 2881 accttacaag acaagatagt gaaagaaaaa tttgctatgg tagcaccaga tgtccaaatt 2941 gaagatggga aaggaacaat cctaatatct tccgaagagg gagagacgga agctaataat 3001 cacaagaagt tgtcagaatt tggaattaga aatggcagcc ggcttcaagc agatgacttc 3061 ctccaggact atactttatt gatcaacatc cttcatagtg aagacctagg aaaggacgtt 3121 gaatttgaag ttgttggtga tgctccggaa aaagtggggc ccaaacaagc tgaagatgct 3181 gccaaaagca taaccaatgg cagtgatgat ggagctcagc cctccacctc cacagctcaa 3241 gagcaagatg acgttctcat agttgattcg gatgaagaag attcttcaaa taatgccgac 3301 gtcagtgaag aagagagaag ccgcaagagg aaattagatg agaaagagaa tctcagtgca 3361 aagaggtcac gtatagaaca gaaggaagag cttgatgatg tcatagcatt agattgagcg 3421 gccgcattaa aaggaggaat taactatggt ggagaaggag gaggctggcg gcggcattag 3481 cgaggaggag gcggcacagt atgaccggca gatccgcctg tggggactgg aggcccagaa 3541 acggctgcgg gcctctcggg tgcttcttgt cggcttgaaa ggacttgggg ctgaaattgc 3601 caagaatctc atcttggcag gagtgaaagg actgaccatg ctggatcacg aacaggtaac 3661 tccagaagat cccggagctc agttcttgat tcgtactggg tctgttggcc gaaatagggc 3721 tgaagcctct ttggagcgag ctcagaatct caaccccatg gtggatgtga aggtggacac 3781 tgaggatata gagaagaaac cagagtcatt tttcactcaa ttcgatgctg tgtgtctgac 3841 ttgctgctcc agggatgtca tagttaaagt tgaccagatc tgtcacaaaa atagcatcaa 3901 gttctttaca ggagatgttt ttggctacca tggatacaca tttgccaatc taggagagca 3961 tgagtttgta gaggagaaaa ctaaagttgc caaagttagc caaggagtag aagatgggcc 4021 cgacaccaag agagcaaaac ttgattcttc tgagacaacg atggtcaaaa agaaggtggt 4081 cttctgccct gttaaagaag ccctggaggt ggactggagc agtgagaaag caaaggctgc 4141 tctgaagcgc acgacctccg actactttct ccttcaagtg ctcttaaagt tccgtacaga 4201 taaaggaaga gatcccagtt ctgatacata tgaggaagat tctgagttgt tgctccagat 4261 acgaaatgat gtgcttgact cactgggtat tagtcctgac ctgcttcctg aggactttgt 4321 caggtactgc ttctccgaga tggccccagt gtgtgcggtg gttggaggga ttttggcaca 4381 ggaaattgtg aaggccctgt ctcagcggga ccctcctcac aacaacttct tcttcttcga 4441 tggcatgaag gggaatggga ttgtggagtg ccttggcccc aagggctagg ctgctgccac 4501 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt 4561 gctgaaacct caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc 4621 ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgaccgg 4681 gtcatcgtgg ccggatcttg cggcccctcg gcttgaacga attgttagac attatttgcc 4741 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg 4801 cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg 4861 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat 4921 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc 4981 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag 5041 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct 5101 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa 5161 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg 5221 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat 5281 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt 5341 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag 5401 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg 5461 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct 5521 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 5581 atacatattt gaatgtattt agaaaaataa acaaatagct agctcactcg gtcgctacgc 5641 tccgggcgtg agactgcggc gggcgctgcg gacacataca aagttaccca cagattccgt 5701 ggataagcag gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc 5761 cataggctcc gccctcctgc cagagttcac ataaacagac gcttttccgg tgcatctgtg 5821 ggagccgtga ggctcaacca tgaatctgac agtacgggcg aaacccgaca ggacttaaag 5881 atccccaccg tttccggcgg gtcgctccct cttgcgctct cctgttccga ccctgccgtt 5941 taccggatac ctgttccgcc tttctccctt acgggaagtg tggcgctttc tcatagctca 6001 cacactggta tctcggctcg gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac 6061 tccccgttca gcccgactgc tgcgccttat ccggtaactg ttcacttgag tccaacccgg 6121 aaaagcacgg taaaacgcca ctggcagcag ccattggtaa ctgggagttc gcagaggatt 6181 tgtttagcta aacacgcggt tgctcttgaa gtgtgcgcca aagtccggct acactggaag 6241 gacagatttg gttgctgtgc tctgcgaaag ccagttacca cggttaagca gttccccaac 6301 tgacttaacc ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga 6361 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct 6421 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata 6481 agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa 6541 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat 6601 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 6661 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt 6721 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca 6781 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg 6841 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac 6901 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg 6961 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac 7021 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga 7081 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta 7141 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt 7201 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg 7261 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt 7321 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt 7381 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag 7441 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc 7501 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca 7561 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt 7621 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg 7681 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac 7741 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac 7801 tcctgcatta ggaaa //