LOCUS       sequence-80812-e        6965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human 6His-tagged SUMO3 and SUMO-E1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6965)
  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 6965)
  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: pSA3
FEATURES             Location/Qualifiers
     source          1..6965
                     /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        531..549
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    550..574
                     /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      3663..3710
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3881..4672)
                     /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(4673..4764)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4812..5550)
                     /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(5760..6842)
                     /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     6804..6823
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6843..6920)
                     /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 ttaagtcgaa cagaaagtaa tcgtattgta cacggccgca taatcgaaat taatacgact
      541 cactataggg gaattgtgag cggataacaa ttccccatct tagtatatta gttaagtata
      601 agaaggagat atacatatgg cagatctcaa ttcccgggtc gacatggcac tgtcgcgggg
      661 gctgccccgg gagctggctg aggcggtggc cgggggccgg gtgctggtgg tgggggcggg
      721 cggcatcggc tgcgagctcc tcaagaatct cgtgctcacc ggtttctccc acatcgacct
      781 gattgatctg gatactattg atgtaagcaa cctcaacaga cagtttttgt ttcaaaagaa
      841 acatgttgga agatcaaagg cacaggttgc caaggaaagt gtactgcagt tttacccgaa
      901 agctaatatc gttgcctacc atgacagcat catgaaccct gactataatg tggaattttt
      961 ccgacagttt atactggtta tgaatgcttt agataacaga gctgcccgaa accatgttaa
     1021 tagaatgtgc ctggcagctg atgttcctct tattgaaagt ggaacagctg ggtatcttgg
     1081 acaagtaact actatcaaaa agggtgtgac cgagtgttat gagtgtcatc ctaagccgac
     1141 ccagagaacc tttcctggct gtacaattcg taacacacct tcagaaccta tacattgcat
     1201 cgtttgggca aagtacttgt tcaaccagtt gtttggggaa gaagatgctg atcaagaagt
     1261 atctcctgac agagctgacc ctgaagctgc ctgggaacca acggaagccg aagccagagc
     1321 tagagcatct aatgaagatg gtgacattaa acgtatttct actaaggaat gggctaaatc
     1381 aactggatat gatccagtta aactttttac caagcttttt aaagatgaca tcaggtatct
     1441 gttgacaatg gacaaactat ggcggaaaag gaaacctcca gttccgttgg actgggctga
     1501 agtacaaagt caaggagaag aaacgaatgc atcagatcaa cagaatgaac cccagttagg
     1561 cctgaaagac cagcaggttc tagatgtaaa gagctatgca cgtctttttt caaagagcat
     1621 cgagactttg agagttcatt tagcagaaaa gggggatgga gctgagctca tatgggataa
     1681 ggatgaccca tctgcaatgg attttgtcac ctctgctgca aacctcagga tgcatatttt
     1741 cagtatgaat atgaagagta gatttgatat caaatcaatg gcagggaaca ttattcctgc
     1801 tattgctact actaatgcag taattgctgg gttgatagta ttggaaggat tgaagatttt
     1861 atcaggaaaa atagaccagt gcagaacaat ttttttgaat aaacaaccaa acccaagaaa
     1921 gaagcttctt gtgccttgtg cactggatcc tcccaacccc aattgttatg tatgtgccag
     1981 caagccagag gtgactgtgc ggctgaatgt ccataaagtg actgttctca ccttacaaga
     2041 caagatagtg aaagaaaaat ttgctatggt agcaccagat gtccaaattg aagatgggaa
     2101 aggaacaatc ctaatatctt ccgaagaggg agagacggaa gctaataatc acaagaagtt
     2161 gtcagaattt ggaattagaa atggcagccg gcttcaagca gatgacttcc tccaggacta
     2221 tactttattg atcaacatcc ttcatagtga agacctagga aaggacgttg aatttgaagt
     2281 tgttggtgat gctccggaaa aagtggggcc caaacaagct gaagatgctg ccaaaagcat
     2341 aaccaatggc agtgatgatg gagctcagcc ctccacctcc acagctcaag agcaagatga
     2401 cgttctcata gttgattcgg atgaagaaga ttcttcaaat aatgccgacg tcagtgaaga
     2461 agagagaagc cgcaagagga aattagatga gaaagagaat ctcagtgcaa agaggtcacg
     2521 tatagaacag aaggaagagc ttgatgatgt catagcatta gattgagcgg ccgcattaaa
     2581 aggaggaatt aactatggtg gagaaggagg aggctggcgg cggcattagc gaggaggagg
     2641 cggcacagta tgaccggcag atccgcctgt ggggactgga ggcccagaaa cggctgcggg
     2701 cctctcgggt gcttcttgtc ggcttgaaag gacttggggc tgaaattgcc aagaatctca
     2761 tcttggcagg agtgaaagga ctgaccatgc tggatcacga acaggtaact ccagaagatc
     2821 ccggagctca gttcttgatt cgtactgggt ctgttggccg aaatagggct gaagcctctt
     2881 tggagcgagc tcagaatctc aaccccatgg tggatgtgaa ggtggacact gaggatatag
     2941 agaagaaacc agagtcattt ttcactcaat tcgatgctgt gtgtctgact tgctgctcca
     3001 gggatgtcat agttaaagtt gaccagatct gtcacaaaaa tagcatcaag ttctttacag
     3061 gagatgtttt tggctaccat ggatacacat ttgccaatct aggagagcat gagtttgtag
     3121 aggagaaaac taaagttgcc aaagttagcc aaggagtaga agatgggccc gacaccaaga
     3181 gagcaaaact tgattcttct gagacaacga tggtcaaaaa gaaggtggtc ttctgccctg
     3241 ttaaagaagc cctggaggtg gactggagca gtgagaaagc aaaggctgct ctgaagcgca
     3301 cgacctccga ctactttctc cttcaagtgc tcttaaagtt ccgtacagat aaaggaagag
     3361 atcccagttc tgatacatat gaggaagatt ctgagttgtt gctccagata cgaaatgatg
     3421 tgcttgactc actgggtatt agtcctgacc tgcttcctga ggactttgtc aggtactgct
     3481 tctccgagat ggccccagtg tgtgcggtgg ttggagggat tttggcacag gaaattgtga
     3541 aggccctgtc tcagcgggac cctcctcaca acaacttctt cttcttcgat ggcatgaagg
     3601 ggaatgggat tgtggagtgc cttggcccca agggctaggc tgctgccacc gctgagcaat
     3661 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaacctc
     3721 aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc
     3781 aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg tcatcgtggc
     3841 cggatcttgc ggcccctcgg cttgaacgaa ttgttagaca ttatttgccg actaccttgg
     3901 tgatctcgcc tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaagc
     3961 gatcttcttc ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg
     4021 ccggcaggcg ctccattgcc cagtcggcag cgacatcctt cggcgcgatt ttgccggtta
     4081 ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt
     4141 cgggcggcga gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag
     4201 gaaccggatc aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg
     4261 cttttgtcag caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa
     4321 gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg
     4381 gaatgatgtc gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc
     4441 caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa
     4501 gccttacggt caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca
     4561 ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc
     4621 caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc atactcttcc
     4681 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4741 aatgtattta gaaaaataaa caaatagcta gctcactcgg tcgctacgct ccgggcgtga
     4801 gactgcggcg ggcgctgcgg acacatacaa agttacccac agattccgtg gataagcagg
     4861 ggactaacat gtgaggcaaa acagcagggc cgcgccggtg gcgtttttcc ataggctccg
     4921 ccctcctgcc agagttcaca taaacagacg cttttccggt gcatctgtgg gagccgtgag
     4981 gctcaaccat gaatctgaca gtacgggcga aacccgacag gacttaaaga tccccaccgt
     5041 ttccggcggg tcgctccctc ttgcgctctc ctgttccgac cctgccgttt accggatacc
     5101 tgttccgcct ttctccctta cgggaagtgt ggcgctttct catagctcac acactggtat
     5161 ctcggctcgg tgtaggtcgt tcgctccaag ctgggctgta agcaagaact ccccgttcag
     5221 cccgactgct gcgccttatc cggtaactgt tcacttgagt ccaacccgga aaagcacggt
     5281 aaaacgccac tggcagcagc cattggtaac tgggagttcg cagaggattt gtttagctaa
     5341 acacgcggtt gctcttgaag tgtgcgccaa agtccggcta cactggaagg acagatttgg
     5401 ttgctgtgct ctgcgaaagc cagttaccac ggttaagcag ttccccaact gacttaacct
     5461 tcgatcaaac cacctcccca ggtggttttt tcgtttacag ggcaaaagat tacgcgcaga
     5521 aaaaaaggat ctcaagaaga tcctttgatc ttttctactg aaccgctcta gatttcagtg
     5581 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc
     5641 tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     5701 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     5761 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5821 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     5881 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     5941 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     6001 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     6061 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     6121 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     6181 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     6241 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     6301 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     6361 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     6421 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     6481 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     6541 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     6601 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     6661 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     6721 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     6781 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     6841 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     6901 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     6961 gaaat
//
