LOCUS       sequence-80814-e        7671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of GST-tagged TARGET proteins fused to SUMO-E2 
            with GyrA intein linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7671)
  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 7671)
  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-IntE2
FEATURES             Location/Qualifiers
     source          1..7671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        72..176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(177..245,246..1037)
                     /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(395..414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(1079..1592)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1274..1295)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1486..1505
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      1703..2291
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2192..2211
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2445..2462
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2484..2503)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2644..2666
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2660..2851)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3408..4490)
                     /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     4452..4471
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4491..4568)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      4721..4807
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4904..4990
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5087..5173
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5270..5356
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5453..5539
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5674..5693
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5674..5692
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5693..5717
                     /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             5732..5754
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5807..6400
                     /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             6917..7573
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(7622..7640)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      join(7636..7671,1..12)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 aggggttttt tgctgaaagg aggaactata tccggataac tacgtcaggt ggcacttttc
       61 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      121 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      181 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      241 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
      301 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
      361 aacgttctcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
      421 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      481 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      541 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      601 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      661 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
      721 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
      781 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
      841 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
      901 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
      961 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1021 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttac
     1081 cccggttgat aatcagaaaa gccccaaaaa caggaagatt gtataagcaa atatttaaat
     1141 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     1201 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaatagc ccgagatagg
     1261 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     1321 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc
     1381 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     1441 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     1501 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     1561 cgccgcgctt aatgcgccgc tacagggcgc gtaaaaggat ctaggtgaag atcctttttg
     1621 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1681 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1741 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1801 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1861 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1921 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1981 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2041 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2101 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2161 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2221 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2281 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2341 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     2401 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     2461 aggaagctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     2521 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     2581 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2641 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     2701 tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca gatgtctgcc tgttcatccg
     2761 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     2821 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     2881 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     2941 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     3001 gggaccagag aaaaatcact cagggtcaat gccagccgaa cgccagcaag acgtagccca
     3061 gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     3121 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     3181 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     3241 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     3301 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     3361 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     3421 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3481 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     3541 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     3601 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     3661 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     3721 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     3781 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     3841 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     3901 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     3961 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     4021 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     4081 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     4141 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     4201 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     4261 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     4321 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     4381 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     4441 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     4501 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     4561 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     4621 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgcc ggcatgccgc
     4681 cctttcgtct tcaagaatta attcccaatt ccccaggcat caaataaaac gaaaggctca
     4741 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     4801 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
     4861 aggacgcccg ccataaactg ccaggaatta attccccagg catcaaataa aacgaaaggc
     4921 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     4981 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     5041 ggcaggacgc ccgccataaa ctgccaggaa ttaattcccc aggcatcaaa taaaacgaaa
     5101 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     5161 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
     5221 gcgggcagga cgcccgccat aaactgccag gaattaattc cccaggcatc aaataaaacg
     5281 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     5341 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     5401 gtggcgggca ggacgcccgc cataaactgc caggaattaa ttccccaggc atcaaataaa
     5461 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     5521 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     5581 agggtggcgg gcaggacgcc cgccataaac tgccaggaat tggggatcgg aattaattcc
     5641 cggtttaaac cggggatctc gatcccgcga aattaatacg actcactata ggggaattgt
     5701 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataac
     5761 catggctagc tcgcgagtcg acggcggccg cgaattcctc gagatgtgca tctgcggaga
     5821 tgcactagtt gccctacccg agggcgagtc ggtacgcatc gccgacatcg tgccgggtgc
     5881 gcggcccaac agtgacaacg ccatcgacct gaaagtcctt gaccggcatg gcaatcccgt
     5941 gctcgccgac cggctgttcc actccggcga gcatccggtg tacacggtgc gtacggtcga
     6001 aggtctgcgt gtgacgggca ccgcgaacca cccgttgttg tgtttggtcg acgtcgccgg
     6061 ggtgccgacc ctgctgtgga agctgatcga cgaaatcaag ccgggcgatt acgcggtgat
     6121 tcaacgcagc gcattcagcg tcgactgtgc aggttttgcc cgcgggaaac ccgaatttgc
     6181 gcccacaacc tacacagtcg gcgtccctgg actggtgcgt ttcttggaag cacaccaccg
     6241 agacccggac gcccaagcta tcgccgacga gctgaccgac gggcggttct actacgcgaa
     6301 agtcgccagt gtcaccgacg ccggcgtgca gccggtgtat agccttcgtg tcgacacggc
     6361 agaccacgcg tttatcacga acgggttcgt cagccacgct actggcctca ccggtctaag
     6421 cttagcatcc atgtcgggga tcgccctcag cagactcgcc caggagagga aagcatggag
     6481 gaaagaccac ccatttggtt tcgtggctgt cccaacaaaa aatcccgatg gcacgatgaa
     6541 cctcatgaac tgggagtgcg ccattccggg aaagaaaggg actccgtggg aaggaggctt
     6601 gtttaaacta cggatgcttt tcaaagatga ttatccatct tcgccaccaa aatgtaaatt
     6661 cgaaccacca ttatttcacc cgaatgtgta cccttcgggg acagtgtgcc tgtccatctt
     6721 agaggaggac aaggactgga ggccagccat cacaatcaaa cagatcctat taggaataca
     6781 ggaacttcta aatgaaccaa atatccaaga cccagctcaa gcagaggcct acacgattta
     6841 ctgccaaaac agagtggagt acgagaaaag ggtccgagca caagccaaga agtttgcgcc
     6901 ctcatcagga tccggaatgt cccctatact aggttattgg aaaattaagg gccttgtgca
     6961 acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg
     7021 cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct
     7081 tccttattat attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat
     7141 agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct
     7201 tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt
     7261 tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga
     7321 tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt
     7381 gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa
     7441 attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc
     7501 cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca
     7561 tcctccaaaa tagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     7621 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt g
//
