LOCUS       sequence-80837-e        8283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human TDG-K330A C-terminally fused to 
            SUMO-E2 and GST.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8283)
  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 8283)
  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: pSC-hTDG-K330A-PreE2
FEATURES             Location/Qualifiers
     source          1..8283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1325..1348
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1856..2512
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(2561..2579)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2575..2622
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2682..2786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2787..2855,2856..3647)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3005..3024)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3689..4202)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3884..3905)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4096..4115
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4313..4901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4802..4821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5055..5072
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5094..5113)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5254..5276
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5270..5461)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6018..7100)
                     /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     7062..7081
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7101..7178)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      7331..7417
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7514..7600
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7697..7783
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7880..7966
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      8063..8149
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataaccat ggaagcggag aacgcgggca gctattccct
      121 tcagcaagct caagcttttt atacgtttcc atttcaacaa ctgatggctg aagctcctaa
      181 tatggcagtt gtgaatgaac agcaaatgcc agaagaagtt ccagccccag ctcctgctca
      241 ggaaccagtg caagaggctc caaaaggaag aaaaagaaaa cccagaacaa cagaaccaaa
      301 acaaccagtg gaacccaaaa aacctgttga gtcaaaaaaa tctggcaagt ctgcaaaatc
      361 aaaagaaaaa caagaaaaaa ttacagacac atttaaagta aaaagaaaag tagaccgttt
      421 taatggtgtt tcagaagctg aacttctgac caagactctc cccgatattt tgaccttcaa
      481 tctggacatt gtcattattg gcataaaccc gggactaatg gctgcttaca aagggcatca
      541 ttaccctgga cctggaaacc atttttggaa gtgtttgttt atgtcagggc tcagtgaggt
      601 ccagctgaac catatggatg atcacactct accagggaag tatggtattg gatttaccaa
      661 catggtggaa aggaccacgc ccggcagcaa agatctctcc agtaaagaat ttcgtgaagg
      721 aggacgtatt ctagtacaga aattacagaa atatcagcca cgaatagcag tgtttaatgg
      781 aaaatgtatt tatgaaattt ttagtaaaga agtttttgga gtaaaggtta agaacttgga
      841 atttgggctt cagccccata agattccaga cacagaaact ctctgctatg ttatgccatc
      901 atccagtgca agatgtgctc agtttcctcg agcccaagac aaagttcatt actacataaa
      961 actgaaggac ttaagagatc agttgaaagg cattgaacga aatatggacg ttcaagaggt
     1021 gcaatataca tttgacctac agcttgccca agaggatgca aagaagatgg ctgttgcgga
     1081 agaaaaatat gatccaggtt atgaggcagc atatggtggt gcttacggag aaaatccatg
     1141 cagcagtgaa ccttgtggct tctcttcaaa tgggctaatt gagagcgtgg agttaagagg
     1201 agaatcagct ttcagtggca ttcctaatgg gcagtggatg acccagtcat ttacagacca
     1261 aattccttcc tttagtaatc actgtggaac acaagaacag gaagaagaaa gccatgctga
     1321 attcctcgag gttctgttcc agggtccagg tggcggaagc ttagcatcca tgtcggggat
     1381 cgccctcagc agactcgccc aggagaggaa agcatggagg aaagaccacc catttggttt
     1441 cgtggctgtc ccaacaaaaa atcccgatgg cacgatgaac ctcatgaact gggagtgcgc
     1501 cattccggga aagaaaggga ctccgtggga aggaggcttg tttaaactac ggatgctttt
     1561 caaagatgat tatccatctt cgccaccaaa atgtaaattc gaaccaccat tatttcaccc
     1621 gaatgtgtac ccttcgggga cagtgtgcct gtccatctta gaggaggaca aggactggag
     1681 gccagccatc acaatcaaac agatcctatt aggaatacag gaacttctaa atgaaccaaa
     1741 tatccaagac ccagctcaag cagaggccta cacgatttac tgccaaaaca gagtggagta
     1801 cgagaaaagg gtccgagcac aagccaagaa gtttgcgccc tcatcaggat ccggaatgtc
     1861 ccctatacta ggttattgga aaattaaggg ccttgtgcaa cccactcgac ttcttttgga
     1921 atatcttgaa gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg ataaatggcg
     1981 aaacaaaaag tttgaattgg gtttggagtt tcccaatctt ccttattata ttgatggtga
     2041 tgttaaatta acacagtcta tggccatcat acgttatata gctgacaagc acaacatgtt
     2101 gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg ttttggatat
     2161 tagatacggt gtttcgagaa ttgcatatag taaagacttt gaaactctca aagttgattt
     2221 tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc ataaaacata
     2281 tttaaatggt gatcatgtaa cccatcctga cttcatgttg tatgacgctc ttgatgttgt
     2341 tttatacatg gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg
     2401 tattgaagct atcccacaaa ttgataagta cttgaaatcc agcaagtata tagcatggcc
     2461 tttgcagggc tggcaagcca cgtttggtgg tggcgaccat cctccaaaat agatccggct
     2521 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     2581 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     2641 tccggataac tacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2701 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2761 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2821 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2881 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2941 gtaagatcct tgagagtttt cgccccgaag aacgttctcc aatgatgagc acttttaaag
     3001 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     3061 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3121 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3181 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3241 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3301 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3361 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3421 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3481 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3541 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3601 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3661 tttactcata tatactttag attgatttac cccggttgat aatcagaaaa gccccaaaaa
     3721 caggaagatt gtataagcaa atatttaaat tgtaaacgtt aatattttgt taaaattcgc
     3781 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3841 ttataaatca aaagaatagc ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3901 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3961 tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc
     4021 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4081 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     4141 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     4201 gtaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4261 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4321 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4381 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4441 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4501 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4561 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4621 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4681 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4741 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4801 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4861 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4921 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4981 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5041 gaacgaccga gcgcagcgag tcagtgagcg aggaagctat ggtgcactct cagtacaatc
     5101 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     5161 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     5221 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     5281 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgca
     5341 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     5401 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     5461 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     5521 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     5581 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     5641 gccagccgaa cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg
     5701 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
     5761 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
     5821 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
     5881 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     5941 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     6001 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6061 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     6121 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     6181 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     6241 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata
     6301 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     6361 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     6421 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     6481 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     6541 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     6601 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     6661 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     6721 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     6781 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     6841 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     6901 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     6961 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     7021 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     7081 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     7141 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     7201 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     7261 cgcaaggaat ggtgcatgcc ggcatgccgc cctttcgtct tcaagaatta attcccaatt
     7321 ccccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     7381 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     7441 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggaatta
     7501 attccccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     7561 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     7621 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggaa
     7681 ttaattcccc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     7741 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     7801 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     7861 gaattaattc cccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     7921 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     7981 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     8041 caggaattaa ttccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     8101 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
     8161 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
     8221 tgccaggaat tggggatcgg aattaattcc cggtttaaac cggggatctc gatcccgcga
     8281 aat
//
