LOCUS       sequence-80805-e        7797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human 6His-tagged SUMO3, SUMO-E1 and 
            SUMO-E2.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2014-07-9"; volume: "9(7):e102157. doi"; pages: " 
            10.1371/journal.pone.0102157. eCollection 2014"
COMMENT     Sequence Label: pSUMO3
FEATURES             Location/Qualifiers
     source          1..7797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
//
