LOCUS       sequence-80803-e        7793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human 6His-tagged SUMO2, SUMO-E1 and 
            SUMO-E2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7793)
  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 7793)
  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: pSUMO2
FEATURES             Location/Qualifiers
     source          1..7793
                     /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..417
                     /codon_start=1
                     /product="human small ubiquitin-related modifier 2"
                     /label=SUMO2
                     /translation="MADEKPKEGVKTENNDHINLKVAGQDGSVVQFKIKRHTPLSKLMK
                     AYCERQGLSMRQIRFRFDGQPINETDTPAQLEMEDEDTIDVFQQQTGG"
     promoter        556..574
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    575..599
                     /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             1131..1175
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      1227..1274
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1359..1377
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1378..1402
                     /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      4491..4538
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4709..5500)
                     /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(5501..5592)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5640..6378)
                     /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(6588..7670)
                     /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     7632..7651
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7671..7748)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagcagcc atcaccatca tcaccacagc
      121 caggatctgg ttccgcgtat ggccgacgaa aagcccaagg aaggagtcaa gactgagaac
      181 aacgatcata ttaatttgaa ggtggcgggg caggatggtt ctgtggtgca gtttaagatt
      241 aagaggcata caccacttag taaactaatg aaagcctatt gtgaacgaca gggattgtca
      301 atgaggcaga tcagattccg atttgacggg caaccaatca atgaaacaga cacacctgca
      361 cagttggaaa tggaggatga agatacaatt gatgtgttcc aacagcagac gggaggttag
      421 gaattcatcg tgactgactg acgctgcagg tcgacaagct tgcggccgca taatgcttaa
      481 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc atgcatcgta ttgtacacgg
      541 ccgcataatc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
      601 catcttagta tattagttaa gtataagaag gagatataca tatgtcgggg atcgccctca
      661 gcagactcgc ccaggagagg aaagcatgga ggaaagacca cccatttggt ttcgtggctg
      721 tcccaacaaa aaatcccgat ggcacgatga acctcatgaa ctgggagtgc gccattccgg
      781 gaaagaaagg gactccgtgg gaaggaggct tgtttaaact acggatgctt ttcaaagatg
      841 attatccatc ttcgccacca aaatgtaaat tcgaaccacc attatttcac ccgaatgtgt
      901 acccttcggg gacagtgtgc ctgtccatct tagaggagga caaggactgg aggccagcca
      961 tcacaatcaa acagatccta ttaggaatac aggaacttct aaatgaacca aatatccaag
     1021 acccagctca agcagaggcc tacacgattt actgccaaaa cagagtggag tacgagaaaa
     1081 gggtccgagc acaagccaag aagtttgcgc cctcataact cgagtctggt aaagaaaccg
     1141 ctgctgcgaa atttgaacgc cagcacatgg actcgtctac tagcgcagct taattaacct
     1201 aggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     1261 tgaggggttt tttgctgaaa cctcaggcat ttgagaagca cggtaccatt aagtcgaaca
     1321 gaaagtaatc gtattgtaca cggccgcata atcgaaatta atacgactca ctatagggga
     1381 attgtgagcg gataacaatt ccccatctta gtatattagt taagtataag aaggagatat
     1441 acatatggca gatctcaatt cccgggtcga catggcactg tcgcgggggc tgccccggga
     1501 gctggctgag gcggtggccg ggggccgggt gctggtggtg ggggcgggcg gcatcggctg
     1561 cgagctcctc aagaatctcg tgctcaccgg tttctcccac atcgacctga ttgatctgga
     1621 tactattgat gtaagcaacc tcaacagaca gtttttgttt caaaagaaac atgttggaag
     1681 atcaaaggca caggttgcca aggaaagtgt actgcagttt tacccgaaag ctaatatcgt
     1741 tgcctaccat gacagcatca tgaaccctga ctataatgtg gaatttttcc gacagtttat
     1801 actggttatg aatgctttag ataacagagc tgcccgaaac catgttaata gaatgtgcct
     1861 ggcagctgat gttcctctta ttgaaagtgg aacagctggg tatcttggac aagtaactac
     1921 tatcaaaaag ggtgtgaccg agtgttatga gtgtcatcct aagccgaccc agagaacctt
     1981 tcctggctgt acaattcgta acacaccttc agaacctata cattgcatcg tttgggcaaa
     2041 gtacttgttc aaccagttgt ttggggaaga agatgctgat caagaagtat ctcctgacag
     2101 agctgaccct gaagctgcct gggaaccaac ggaagccgaa gccagagcta gagcatctaa
     2161 tgaagatggt gacattaaac gtatttctac taaggaatgg gctaaatcaa ctggatatga
     2221 tccagttaaa ctttttacca agctttttaa agatgacatc aggtatctgt tgacaatgga
     2281 caaactatgg cggaaaagga aacctccagt tccgttggac tgggctgaag tacaaagtca
     2341 aggagaagaa acgaatgcat cagatcaaca gaatgaaccc cagttaggcc tgaaagacca
     2401 gcaggttcta gatgtaaaga gctatgcacg tcttttttca aagagcatcg agactttgag
     2461 agttcattta gcagaaaagg gggatggagc tgagctcata tgggataagg atgacccatc
     2521 tgcaatggat tttgtcacct ctgctgcaaa cctcaggatg catattttca gtatgaatat
     2581 gaagagtaga tttgatatca aatcaatggc agggaacatt attcctgcta ttgctactac
     2641 taatgcagta attgctgggt tgatagtatt ggaaggattg aagattttat caggaaaaat
     2701 agaccagtgc agaacaattt ttttgaataa acaaccaaac ccaagaaaga agcttcttgt
     2761 gccttgtgca ctggatcctc ccaaccccaa ttgttatgta tgtgccagca agccagaggt
     2821 gactgtgcgg ctgaatgtcc ataaagtgac tgttctcacc ttacaagaca agatagtgaa
     2881 agaaaaattt gctatggtag caccagatgt ccaaattgaa gatgggaaag gaacaatcct
     2941 aatatcttcc gaagagggag agacggaagc taataatcac aagaagttgt cagaatttgg
     3001 aattagaaat ggcagccggc ttcaagcaga tgacttcctc caggactata ctttattgat
     3061 caacatcctt catagtgaag acctaggaaa ggacgttgaa tttgaagttg ttggtgatgc
     3121 tccggaaaaa gtggggccca aacaagctga agatgctgcc aaaagcataa ccaatggcag
     3181 tgatgatgga gctcagccct ccacctccac agctcaagag caagatgacg ttctcatagt
     3241 tgattcggat gaagaagatt cttcaaataa tgccgacgtc agtgaagaag agagaagccg
     3301 caagaggaaa ttagatgaga aagagaatct cagtgcaaag aggtcacgta tagaacagaa
     3361 ggaagagctt gatgatgtca tagcattaga ttgagcggcc gcattaaaag gaggaattaa
     3421 ctatggtgga gaaggaggag gctggcggcg gcattagcga ggaggaggcg gcacagtatg
     3481 accggcagat ccgcctgtgg ggactggagg cccagaaacg gctgcgggcc tctcgggtgc
     3541 ttcttgtcgg cttgaaagga cttggggctg aaattgccaa gaatctcatc ttggcaggag
     3601 tgaaaggact gaccatgctg gatcacgaac aggtaactcc agaagatccc ggagctcagt
     3661 tcttgattcg tactgggtct gttggccgaa atagggctga agcctctttg gagcgagctc
     3721 agaatctcaa ccccatggtg gatgtgaagg tggacactga ggatatagag aagaaaccag
     3781 agtcattttt cactcaattc gatgctgtgt gtctgacttg ctgctccagg gatgtcatag
     3841 ttaaagttga ccagatctgt cacaaaaata gcatcaagtt ctttacagga gatgtttttg
     3901 gctaccatgg atacacattt gccaatctag gagagcatga gtttgtagag gagaaaacta
     3961 aagttgccaa agttagccaa ggagtagaag atgggcccga caccaagaga gcaaaacttg
     4021 attcttctga gacaacgatg gtcaaaaaga aggtggtctt ctgccctgtt aaagaagccc
     4081 tggaggtgga ctggagcagt gagaaagcaa aggctgctct gaagcgcacg acctccgact
     4141 actttctcct tcaagtgctc ttaaagttcc gtacagataa aggaagagat cccagttctg
     4201 atacatatga ggaagattct gagttgttgc tccagatacg aaatgatgtg cttgactcac
     4261 tgggtattag tcctgacctg cttcctgagg actttgtcag gtactgcttc tccgagatgg
     4321 ccccagtgtg tgcggtggtt ggagggattt tggcacagga aattgtgaag gccctgtctc
     4381 agcgggaccc tcctcacaac aacttcttct tcttcgatgg catgaagggg aatgggattg
     4441 tggagtgcct tggccccaag ggctaggctg ctgccaccgc tgagcaataa ctagcataac
     4501 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag gcatttgaga
     4561 agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag
     4621 cggctattta acgaccctgc cctgaaccga cgaccgggtc atcgtggccg gatcttgcgg
     4681 cccctcggct tgaacgaatt gttagacatt atttgccgac taccttggtg atctcgcctt
     4741 tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt
     4801 gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct
     4861 ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc
     4921 aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt
     4981 tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa
     5041 agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca
     5101 agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc
     5161 gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt
     5221 cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg
     5281 aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca
     5341 ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt
     5401 acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg
     5461 atagttgagt cgatacttcg gcgatcaccg cttccctcat actcttcctt tttcaatatt
     5521 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5581 aaaataaaca aatagctagc tcactcggtc gctacgctcc gggcgtgaga ctgcggcggg
     5641 cgctgcggac acatacaaag ttacccacag attccgtgga taagcagggg actaacatgt
     5701 gaggcaaaac agcagggccg cgccggtggc gtttttccat aggctccgcc ctcctgccag
     5761 agttcacata aacagacgct tttccggtgc atctgtggga gccgtgaggc tcaaccatga
     5821 atctgacagt acgggcgaaa cccgacagga cttaaagatc cccaccgttt ccggcgggtc
     5881 gctccctctt gcgctctcct gttccgaccc tgccgtttac cggatacctg ttccgccttt
     5941 ctcccttacg ggaagtgtgg cgctttctca tagctcacac actggtatct cggctcggtg
     6001 taggtcgttc gctccaagct gggctgtaag caagaactcc ccgttcagcc cgactgctgc
     6061 gccttatccg gtaactgttc acttgagtcc aacccggaaa agcacggtaa aacgccactg
     6121 gcagcagcca ttggtaactg ggagttcgca gaggatttgt ttagctaaac acgcggttgc
     6181 tcttgaagtg tgcgccaaag tccggctaca ctggaaggac agatttggtt gctgtgctct
     6241 gcgaaagcca gttaccacgg ttaagcagtt ccccaactga cttaaccttc gatcaaacca
     6301 cctccccagg tggttttttc gtttacaggg caaaagatta cgcgcagaaa aaaaggatct
     6361 caagaagatc ctttgatctt ttctactgaa ccgctctaga tttcagtgca atttatctct
     6421 tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc atgttagtca
     6481 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     6541 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     6601 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6661 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     6721 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     6781 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     6841 tcggtatcgt cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta
     6901 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     6961 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     7021 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     7081 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     7141 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     7201 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     7261 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     7321 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     7381 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     7441 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     7501 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     7561 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     7621 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     7681 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     7741 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga aat
//
