LOCUS       sequence-80849-e        8394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human TDG-K330A C-terminally fused to GyrA 
            intein and GST.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8394)
  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 8394)
  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: pTXG-hTDG-K330A
FEATURES             Location/Qualifiers
     source          1..8394
                     /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             1334..1927
                     /codon_start=1
                     /gene="Mycobacterium xenopi gyrA"
                     /product="modified GyrA intein (Southworth et al., 1999)"
                     /label=Mxe GyrA intein
                     /note="N-terminal cleavage of this intein can be induced by
                     thiols."
                     /translation="CICGDALVALPEGESVRIADIVPGARPNSDNAIDLKVLDRHGNPV
                     LADRLFHSGEHPVYTVRTVEGLRVTGTANHPLLCLVDVAGVPTLLWKLIDEIKPGDYAV
                     IQRSAFSVDCAGFARGKPEFAPTTYTVGVPGLVRFLEAHHRDPDAQAIADELTDGRFYY
                     AKVASVTDAGVQPVYSLRVDTADHAFITNGFVSHA"
     CDS             1967..2623
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(2672..2690)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2686..2733
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2793..2897
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2898..2966,2967..3758)
                     /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(3116..3135)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3800..4313)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3995..4016)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4207..4226
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4424..5012
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4913..4932
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5166..5183
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5205..5224)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5365..5387
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5381..5572)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6129..7211)
                     /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     7173..7192
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7212..7289)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      7442..7528
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7625..7711
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7808..7894
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7991..8077
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      8174..8260
                     /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 atgtgcatct gcggagatgc actagttgcc ctacccgagg gcgagtcggt
     1381 acgcatcgcc gacatcgtgc cgggtgcgcg gcccaacagt gacaacgcca tcgacctgaa
     1441 agtccttgac cggcatggca atcccgtgct cgccgaccgg ctgttccact ccggcgagca
     1501 tccggtgtac acggtgcgta cggtcgaagg tctgcgtgtg acgggcaccg cgaaccaccc
     1561 gttgttgtgt ttggtcgacg tcgccggggt gccgaccctg ctgtggaagc tgatcgacga
     1621 aatcaagccg ggcgattacg cggtgattca acgcagcgca ttcagcgtcg actgtgcagg
     1681 ttttgcccgc gggaaacccg aatttgcgcc cacaacctac acagtcggcg tccctggact
     1741 ggtgcgtttc ttggaagcac accaccgaga cccggacgcc caagctatcg ccgacgagct
     1801 gaccgacggg cggttctact acgcgaaagt cgccagtgtc accgacgccg gcgtgcagcc
     1861 ggtgtatagc cttcgtgtcg acacggcaga ccacgcgttt atcacgaacg ggttcgtcag
     1921 ccacgctact ggcctcaccg gtctgaactc aggcctcgga tccggaatgt cccctatact
     1981 aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga
     2041 agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa
     2101 gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt
     2161 aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg
     2221 tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg
     2281 tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa
     2341 gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg
     2401 tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat
     2461 ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc
     2521 tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg
     2581 ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa tagatccggc tgctaacaaa
     2641 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     2701 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggataa
     2761 ctacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2821 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2881 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2941 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3001 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3061 ttgagagttt tcgccccgaa gaacgttctc caatgatgag cacttttaaa gttctgctat
     3121 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     3181 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3241 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3301 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3361 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3421 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3481 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3541 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3601 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3661 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3721 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3781 atatacttta gattgattta ccccggttga taatcagaaa agccccaaaa acaggaagat
     3841 tgtataagca aatatttaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt
     3901 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3961 aaaagaatag cccgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     4021 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4081 acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4141 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4201 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4261 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtaaaagga
     4321 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4381 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4441 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4501 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4561 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4621 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4681 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4741 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4801 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4861 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4921 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4981 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5041 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5101 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5161 agcgcagcga gtcagtgagc gaggaagcta tggtgcactc tcagtacaat ctgctctgat
     5221 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     5281 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     5341 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     5401 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtgc agcgattcac
     5461 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     5521 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     5581 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     5641 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     5701 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagccga
     5761 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct
     5821 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     5881 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     5941 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     6001 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     6061 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     6121 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6181 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     6241 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     6301 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     6361 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     6421 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     6481 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     6541 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     6601 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     6661 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     6721 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     6781 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     6841 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     6901 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     6961 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     7021 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     7081 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     7141 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     7201 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     7261 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     7321 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     7381 tggtgcatgc cggcatgccg ccctttcgtc ttcaagaatt aattcccaat tccccaggca
     7441 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     7501 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     7561 acggcccgga gggtggcggg caggacgccc gccataaact gccaggaatt aattccccag
     7621 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     7681 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
     7741 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccagga attaattccc
     7801 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     7861 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     7921 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggaattaatt
     7981 ccccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     8041 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     8101 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggaatta
     8161 attccccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     8221 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     8281 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggaa
     8341 ttggggatcg gaattaattc ccggtttaaa ccggggatct cgatcccgcg aaat
//
