LOCUS       sequence-80844-e        7194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of TARGET proteins C-terminally fused to GyrA 
            intein and GST.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7194)
  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 7194)
  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
FEATURES             Location/Qualifiers
     source          1..7194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..134
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(135..203,204..995)
                     /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(353..372)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(1037..1550)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1232..1253)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1444..1463
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      1661..2249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2150..2169
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2403..2420
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2442..2461)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2602..2624
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2618..2809)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3366..4448)
                     /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     4410..4429
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4449..4526)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      4679..4765
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4862..4948
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5045..5131
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5228..5314
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5411..5497
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5632..5651
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5632..5650
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5651..5675
                     /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             5690..5712
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5765..6358
                     /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             6398..7054
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(7103..7121)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7117..7164
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
       61 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      121 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
      181 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
      241 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
      301 agagttttcg ccccgaagaa cgttctccaa tgatgagcac ttttaaagtt ctgctatgtg
      361 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
      421 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
      481 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
      541 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
      601 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
      661 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
      721 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
      781 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
      841 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
      901 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
      961 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1021 tactttagat tgatttaccc cggttgataa tcagaaaagc cccaaaaaca ggaagattgt
     1081 ataagcaaat atttaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     1141 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     1201 agaatagccc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     1261 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     1321 tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     1381 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     1441 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     1501 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt aaaaggatct
     1561 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1621 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1681 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1741 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1801 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1861 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     1921 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     1981 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2041 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2101 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2161 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2221 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2281 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2341 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2401 gcagcgagtc agtgagcgag gaagctatgg tgcactctca gtacaatctg ctctgatgcc
     2461 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2521 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2581 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2641 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgcagc gattcacaga
     2701 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2761 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2821 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2881 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     2941 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagccgaacg
     3001 ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc tgcttctcgc
     3061 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     3121 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     3181 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     3241 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     3301 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     3361 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3421 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     3481 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     3541 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     3601 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     3661 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     3721 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     3781 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     3841 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     3901 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     3961 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     4021 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     4081 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     4141 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     4201 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     4261 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     4321 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     4381 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     4441 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     4501 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     4561 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     4621 tgcatgccgg catgccgccc tttcgtcttc aagaattaat tcccaattcc ccaggcatca
     4681 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     4741 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
     4801 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggaattaat tccccaggca
     4861 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     4921 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     4981 acggcccgga gggtggcggg caggacgccc gccataaact gccaggaatt aattccccag
     5041 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     5101 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
     5161 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccagga attaattccc
     5221 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     5281 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     5341 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggaattaatt
     5401 ccccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     5461 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     5521 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggaattg
     5581 gggatcggaa ttaattcccg gtttaaaccg gggatctcga tcccgcgaaa ttaatacgac
     5641 tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact
     5701 ttaagaagga gatataacca tggctagctc gcgagtcgac ggcggccgcg aattcctcga
     5761 gatgtgcatc tgcggagatg cactagttgc cctacccgag ggcgagtcgg tacgcatcgc
     5821 cgacatcgtg ccgggtgcgc ggcccaacag tgacaacgcc atcgacctga aagtccttga
     5881 ccggcatggc aatcccgtgc tcgccgaccg gctgttccac tccggcgagc atccggtgta
     5941 cacggtgcgt acggtcgaag gtctgcgtgt gacgggcacc gcgaaccacc cgttgttgtg
     6001 tttggtcgac gtcgccgggg tgccgaccct gctgtggaag ctgatcgacg aaatcaagcc
     6061 gggcgattac gcggtgattc aacgcagcgc attcagcgtc gactgtgcag gttttgcccg
     6121 cgggaaaccc gaatttgcgc ccacaaccta cacagtcggc gtccctggac tggtgcgttt
     6181 cttggaagca caccaccgag acccggacgc ccaagctatc gccgacgagc tgaccgacgg
     6241 gcggttctac tacgcgaaag tcgccagtgt caccgacgcc ggcgtgcagc cggtgtatag
     6301 ccttcgtgtc gacacggcag accacgcgtt tatcacgaac gggttcgtca gccacgctac
     6361 tggcctcacc ggtctgaact caggcctcgg atccggaatg tcccctatac taggttattg
     6421 gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg aagaaaaata
     6481 tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt
     6541 gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat taacacagtc
     6601 tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga
     6661 gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg gtgtttcgag
     6721 aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca agctacctga
     6781 aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg gtgatcatgt
     6841 aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca tggacccaat
     6901 gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag ctatcccaca
     6961 aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg gctggcaagc
     7021 cacgtttggt ggtggcgacc atcctccaaa atagatccgg ctgctaacaa agcccgaaag
     7081 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     7141 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata acta
//
