LOCUS       sequence-80839-e        6879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human XRCC1-K176R with N-terminal GST-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6879)
  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 6879)
  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: pGEX-hXRCC1-K176R
FEATURES             Location/Qualifiers
     source          1..6879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    36..52
                     /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             75..728
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     686..708
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             735..758
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2746..2768)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2913..2931)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2999..3103
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3104..3172,3173..3964)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3322..3341)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4135..4723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4624..4643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4877..4894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4967..5044
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5045..6127
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5064..5083)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6176..6181,6182..6199,6200..6206)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6214..6230
                     /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)."
     primer_bind     6238..6254
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6238..6254
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6250..6777
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6270..6287)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6270..6286)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6279..6301)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6879,1..5,6..21,22..28)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 tgacaattaa tcatcggctc gtataatgtg tggaattgtg agcggataac aatttcacac
       61 aggaaacagt attcatgtcc cctatactag gttattggaa aattaagggc cttgtgcaac
      121 ccactcgact tcttttggaa tatcttgaag aaaaatatga agagcatttg tatgagcgcg
      181 atgaaggtga taaatggcga aacaaaaagt ttgaattggg tttggagttt cccaatcttc
      241 cttattatat tgatggtgat gttaaattaa cacagtctat ggccatcata cgttatatag
      301 ctgacaagca caacatgttg ggtggttgtc caaaagagcg tgcagagatt tcaatgcttg
      361 aaggagcggt tttggatatt agatacggtg tttcgagaat tgcatatagt aaagactttg
      421 aaactctcaa agttgatttt cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc
      481 gtttatgtca taaaacatat ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt
      541 atgacgctct tgatgttgtt ttatacatgg acccaatgtg cctggatgcg ttcccaaaat
      601 tagtttgttt taaaaaacgt attgaagcta tcccacaaat tgataagtac ttgaaatcca
      661 gcaagtatat agcatggcct ttgcagggct ggcaagccac gtttggtggt ggcgaccatc
      721 ctccaaaatc ggatctggaa gttctgttcc aggggcccct gggatccccg aattctccgg
      781 agatccgcct ccgccatgtc gtgtcctgca gcagccagga ctcgactcac tgtgcagaaa
      841 atcttctcaa ggcagacact taccgaaaat ggcgggcagc caaggcaggc gagaagacca
      901 tctctgtggt cctacagttg gagaaggagg agcagataca cagtgtggac attgggaatg
      961 atggctcagc tttcgtggag gtgctggtgg gcagttcagc tggaggcgct ggggagcaag
     1021 actatgaggt ccttctggtc acctcatctt tcatgtcccc ttccgagagc cgcagtggct
     1081 caaaccccaa ccgcgttcgc atgtttgggc ctgacaagct ggtccgggca gccgccgaga
     1141 agcgctggga ccgggtcaaa attgtttgca gccagcccta cagcaaggac tccccctttg
     1201 gcttgagttt tgtacggttt catagccccc cagacaaaga tgaggcagag gccccgtccc
     1261 agaaggtgac agtgaccaag cttggccagt tccgtgtgag ggaggaggat gagagcgcca
     1321 actctctgag gccgggggct ctcttcttca gccggatcaa caagacatcc ccagtcacag
     1381 ccagcgaccc ggcaggacct agctatgcag ctgctaccct ccaggcttct agtgctgcct
     1441 cctcagcctc tccagtctcc agggccatag gcagcacctc caagccccag gagtctccca
     1501 aagggaagag gaagttggat ttgaaccaag aagaaaagaa gacccccagc aaaccaccag
     1561 cccagctgtc gccatctgtt cccaagagac ctaaattgcc agctccaact cgtaccccag
     1621 ccacagcccc agtccctgcc cgagcacagg gggcagtgac aggcaaaccc cgaggagaag
     1681 gcaccgagcc cagacgaccc cgagctggcc cagaggagct ggggaagatc cttcagggtg
     1741 tggtagtggt gctgagtggc ttccagaacc ccttccgctc cgagctgcga gataaggccc
     1801 tagagcttgg ggccaagtat cggccagact ggacccggga cagcacgcac ctcatctgtg
     1861 cctttgccaa cacccccaag tacagccagg tcctaggcct gggaggccgc atcgtgcgta
     1921 aggagtgggt gctggactgt caccgcatgc gtcggcggct gccctcccgg aggtacctca
     1981 tggcagggcc aggttccagc agtgaggagg atgaggcctc tcacagcggt ggcagcggag
     2041 atgaagcccc caagcttcct cagaagcaac cccagaccaa aaccaagccc actcaggcag
     2101 ctggacccag ctcaccccag aagcccccaa cccctgaaga gaccaaagca gcctcaccag
     2161 tgctccagga agatatagac attgaggggg tacagtcaga aggacaggac aatggggcgg
     2221 aagattctgg ggacacagag gatgagctga ggagggtggc agagcagaag gaacacagac
     2281 tgccccctgg ccaggaggag aatggggaag acccgtatgc aggctccacg gatgagaaca
     2341 cggacagtga ggaacaccag gagcctcctg atctgccagt ccctgagctc ccagatttct
     2401 tccagggcaa gcacttcttt ctttacgggg agttccctgg ggacgagcgg cggaaactca
     2461 tccgatacgt cacagccttc aatggggagc tcgaggacta tatgagtgac cgggttcagt
     2521 ttgtgatcac agcacaggaa tgggatccca gctttgagga ggccctgatg gacaacccct
     2581 ccctggcatt cgttcgtccc cgatggatct acagttgcaa tgagaagcag aagttacttc
     2641 ctcaccagct ctatggggtg gtgccgcaag cctaggtcga ctcgagcggc cgcatcgtga
     2701 ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     2761 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     2821 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     2881 tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     2941 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3001 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3061 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3121 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3181 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3241 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3301 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     3361 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3421 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3481 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3541 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3601 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     3661 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3721 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3781 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3841 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3901 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3961 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4021 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4081 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4141 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4201 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4261 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     4321 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4381 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4441 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4501 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4561 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4621 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4681 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4741 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4801 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4861 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     4921 ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg
     4981 gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     5041 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
     5101 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
     5161 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
     5221 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
     5281 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
     5341 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
     5401 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
     5461 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
     5521 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
     5581 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
     5641 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
     5701 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
     5761 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
     5821 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag
     5881 ctcatgttat atcccgccgt caaccaccat caaacaggat tttcgcctgc tggggcaaac
     5941 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     6001 gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc
     6061 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     6121 gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccc caggctttac
     6181 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     6241 gaaacagcta tgaccatgat tacggattca ctggccgtcg ttttacaacg tcgtgactgg
     6301 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     6361 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     6421 gaatggcgct ttgcctggtt tccggcacca gaagcggtgc cggaaagctg gctggagtgc
     6481 gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat
     6541 gcgcccatct acaccaacgt gacctatccc attacggtca atccgccgtt tgttcccacg
     6601 gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg gctacaggaa
     6661 ggccagacgc gaattatttt tgatggcgtt ggaattagct tatcgactgc acggtgcacc
     6721 aatgcttctg gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc
     6781 actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataatgttt tttgcgccga
     6841 catcataacg gttctggcaa atattctgaa atgagctgt
//
