LOCUS       sequence-80811-e        6961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human 6His-tagged SUMO2 and SUMO-E1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6961)
  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 6961)
  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: pSA2
FEATURES             Location/Qualifiers
     source          1..6961
                     /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        527..545
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    546..570
                     /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)."
     primer_bind     complement(3645..3663)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3659..3706
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3877..4668)
                     /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(4669..4760)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4808..5546)
                     /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(5756..6838)
                     /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     6800..6819
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6839..6916)
                     /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 gtcgaacaga aagtaatcgt attgtacacg gccgcataat cgaaattaat acgactcact
      541 ataggggaat tgtgagcgga taacaattcc ccatcttagt atattagtta agtataagaa
      601 ggagatatac atatggcaga tctcaattcc cgggtcgaca tggcactgtc gcgggggctg
      661 ccccgggagc tggctgaggc ggtggccggg ggccgggtgc tggtggtggg ggcgggcggc
      721 atcggctgcg agctcctcaa gaatctcgtg ctcaccggtt tctcccacat cgacctgatt
      781 gatctggata ctattgatgt aagcaacctc aacagacagt ttttgtttca aaagaaacat
      841 gttggaagat caaaggcaca ggttgccaag gaaagtgtac tgcagtttta cccgaaagct
      901 aatatcgttg cctaccatga cagcatcatg aaccctgact ataatgtgga atttttccga
      961 cagtttatac tggttatgaa tgctttagat aacagagctg cccgaaacca tgttaataga
     1021 atgtgcctgg cagctgatgt tcctcttatt gaaagtggaa cagctgggta tcttggacaa
     1081 gtaactacta tcaaaaaggg tgtgaccgag tgttatgagt gtcatcctaa gccgacccag
     1141 agaacctttc ctggctgtac aattcgtaac acaccttcag aacctataca ttgcatcgtt
     1201 tgggcaaagt acttgttcaa ccagttgttt ggggaagaag atgctgatca agaagtatct
     1261 cctgacagag ctgaccctga agctgcctgg gaaccaacgg aagccgaagc cagagctaga
     1321 gcatctaatg aagatggtga cattaaacgt atttctacta aggaatgggc taaatcaact
     1381 ggatatgatc cagttaaact ttttaccaag ctttttaaag atgacatcag gtatctgttg
     1441 acaatggaca aactatggcg gaaaaggaaa cctccagttc cgttggactg ggctgaagta
     1501 caaagtcaag gagaagaaac gaatgcatca gatcaacaga atgaacccca gttaggcctg
     1561 aaagaccagc aggttctaga tgtaaagagc tatgcacgtc ttttttcaaa gagcatcgag
     1621 actttgagag ttcatttagc agaaaagggg gatggagctg agctcatatg ggataaggat
     1681 gacccatctg caatggattt tgtcacctct gctgcaaacc tcaggatgca tattttcagt
     1741 atgaatatga agagtagatt tgatatcaaa tcaatggcag ggaacattat tcctgctatt
     1801 gctactacta atgcagtaat tgctgggttg atagtattgg aaggattgaa gattttatca
     1861 ggaaaaatag accagtgcag aacaattttt ttgaataaac aaccaaaccc aagaaagaag
     1921 cttcttgtgc cttgtgcact ggatcctccc aaccccaatt gttatgtatg tgccagcaag
     1981 ccagaggtga ctgtgcggct gaatgtccat aaagtgactg ttctcacctt acaagacaag
     2041 atagtgaaag aaaaatttgc tatggtagca ccagatgtcc aaattgaaga tgggaaagga
     2101 acaatcctaa tatcttccga agagggagag acggaagcta ataatcacaa gaagttgtca
     2161 gaatttggaa ttagaaatgg cagccggctt caagcagatg acttcctcca ggactatact
     2221 ttattgatca acatccttca tagtgaagac ctaggaaagg acgttgaatt tgaagttgtt
     2281 ggtgatgctc cggaaaaagt ggggcccaaa caagctgaag atgctgccaa aagcataacc
     2341 aatggcagtg atgatggagc tcagccctcc acctccacag ctcaagagca agatgacgtt
     2401 ctcatagttg attcggatga agaagattct tcaaataatg ccgacgtcag tgaagaagag
     2461 agaagccgca agaggaaatt agatgagaaa gagaatctca gtgcaaagag gtcacgtata
     2521 gaacagaagg aagagcttga tgatgtcata gcattagatt gagcggccgc attaaaagga
     2581 ggaattaact atggtggaga aggaggaggc tggcggcggc attagcgagg aggaggcggc
     2641 acagtatgac cggcagatcc gcctgtgggg actggaggcc cagaaacggc tgcgggcctc
     2701 tcgggtgctt cttgtcggct tgaaaggact tggggctgaa attgccaaga atctcatctt
     2761 ggcaggagtg aaaggactga ccatgctgga tcacgaacag gtaactccag aagatcccgg
     2821 agctcagttc ttgattcgta ctgggtctgt tggccgaaat agggctgaag cctctttgga
     2881 gcgagctcag aatctcaacc ccatggtgga tgtgaaggtg gacactgagg atatagagaa
     2941 gaaaccagag tcatttttca ctcaattcga tgctgtgtgt ctgacttgct gctccaggga
     3001 tgtcatagtt aaagttgacc agatctgtca caaaaatagc atcaagttct ttacaggaga
     3061 tgtttttggc taccatggat acacatttgc caatctagga gagcatgagt ttgtagagga
     3121 gaaaactaaa gttgccaaag ttagccaagg agtagaagat gggcccgaca ccaagagagc
     3181 aaaacttgat tcttctgaga caacgatggt caaaaagaag gtggtcttct gccctgttaa
     3241 agaagccctg gaggtggact ggagcagtga gaaagcaaag gctgctctga agcgcacgac
     3301 ctccgactac tttctccttc aagtgctctt aaagttccgt acagataaag gaagagatcc
     3361 cagttctgat acatatgagg aagattctga gttgttgctc cagatacgaa atgatgtgct
     3421 tgactcactg ggtattagtc ctgacctgct tcctgaggac tttgtcaggt actgcttctc
     3481 cgagatggcc ccagtgtgtg cggtggttgg agggattttg gcacaggaaa ttgtgaaggc
     3541 cctgtctcag cgggaccctc ctcacaacaa cttcttcttc ttcgatggca tgaaggggaa
     3601 tgggattgtg gagtgccttg gccccaaggg ctaggctgct gccaccgctg agcaataact
     3661 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aacctcaggc
     3721 atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata
     3781 gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcat cgtggccgga
     3841 tcttgcggcc cctcggcttg aacgaattgt tagacattat ttgccgacta ccttggtgat
     3901 ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc
     3961 ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg
     4021 caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc
     4081 gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg
     4141 cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac
     4201 cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt
     4261 tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat
     4321 gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat
     4381 gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg
     4441 ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct
     4501 tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc
     4561 ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac
     4621 tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcatac tcttcctttt
     4681 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4741 tatttagaaa aataaacaaa tagctagctc actcggtcgc tacgctccgg gcgtgagact
     4801 gcggcgggcg ctgcggacac atacaaagtt acccacagat tccgtggata agcaggggac
     4861 taacatgtga ggcaaaacag cagggccgcg ccggtggcgt ttttccatag gctccgccct
     4921 cctgccagag ttcacataaa cagacgcttt tccggtgcat ctgtgggagc cgtgaggctc
     4981 aaccatgaat ctgacagtac gggcgaaacc cgacaggact taaagatccc caccgtttcc
     5041 ggcgggtcgc tccctcttgc gctctcctgt tccgaccctg ccgtttaccg gatacctgtt
     5101 ccgcctttct cccttacggg aagtgtggcg ctttctcata gctcacacac tggtatctcg
     5161 gctcggtgta ggtcgttcgc tccaagctgg gctgtaagca agaactcccc gttcagcccg
     5221 actgctgcgc cttatccggt aactgttcac ttgagtccaa cccggaaaag cacggtaaaa
     5281 cgccactggc agcagccatt ggtaactggg agttcgcaga ggatttgttt agctaaacac
     5341 gcggttgctc ttgaagtgtg cgccaaagtc cggctacact ggaaggacag atttggttgc
     5401 tgtgctctgc gaaagccagt taccacggtt aagcagttcc ccaactgact taaccttcga
     5461 tcaaaccacc tccccaggtg gttttttcgt ttacagggca aaagattacg cgcagaaaaa
     5521 aaggatctca agaagatcct ttgatctttt ctactgaacc gctctagatt tcagtgcaat
     5581 ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat
     5641 gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     5701 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     5761 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5821 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     5881 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     5941 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     6001 agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg
     6061 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     6121 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     6181 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     6241 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     6301 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     6361 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     6421 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     6481 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     6541 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     6601 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     6661 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     6721 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     6781 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     6841 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     6901 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa
     6961 t
//
