LOCUS       sequence-408528-        7137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of PAR-deficient hXRCC1pd with 6xHis tag, under
            the control of T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7137)
  AUTHORS   Schwarz SD, Xu J, Gunasekera K, Schurmann D, Vagbo CB, Ferrari E, 
            Slupphaug G, Hottiger MO, Schar P, Steinacher R
  TITLE     Covalent PARylation of DNA base excision repair proteins regulates 
            DNA demethylation.
  JOURNAL   Nat Commun. 2024 Jan 2;15(1):184. doi: 10.1038/s41467-023-44209-8.
  PUBMED    38167803
REFERENCE   2  (bases 1 to 7137)
  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: "Nat 
            Commun."; date: "2024-01-2"; volume: "15(1):184. doi"; pages: " 
            10.1038/s41467-023-44209-8"
COMMENT     Sequence Label: pET28-hXRCC1pd-HIS
FEATURES             Location/Qualifiers
     source          1..7137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     44..63
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     147..166
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        147..165
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    166..190
                     /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             205..227
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2148..2165
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2218..2236)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2232..2279
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2316..2771
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2403..2422)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2613..2634
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2863..3678)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2863..3678)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3586..3605
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3670..3689
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3800..4388
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4289..4308
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4542..4559
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4574..4713
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4639..4658)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4799..4821
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4815..5006)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5815..6897)
                     /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     6859..6878
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6898..6975)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
       61 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
      121 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
      181 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggat
      241 ccatgccgga gatccgcctc cgccatgtcg tgtcctgcag cagccaggac tcgactcact
      301 gtgcagaaaa tcttctcaag gcagacactt accgaaaatg gcgggcagcc aaggcaggcg
      361 agaagaccat ctctgtggtc ctacagttgg agaaggagga gcagatacac agtgtggaca
      421 ttgggaatga tggctcagct ttcgtggagg tgctggtggg cagttcagct ggaggcgctg
      481 gggagcaaga ctatgaggtc cttctggtca cctcatcttt catgtcccct tccgagagcc
      541 gcagtggcgc taaccccaac cgcgttcgca tgtttgggcc tgacaagctg gtccgggcag
      601 ccgccgagaa gcgctgggac cgggtcaaaa ttgtttgcag ccagccctac agcaaggact
      661 ccccctttgg cttgagtttt gtacggtttc atagcccccc agacaaagat gaggcagagg
      721 ccccgtccca gaaggtgaca gtgaccaagc ttggccagtt ccgtgtgaag gaggaggatg
      781 agagcgccaa cgctctggct ccgggggctc tcttcttcgc tcggatcaac aagacatccc
      841 cagtcacagc cagcgacccg gcaggaccta gctatgcagc tgctaccctc caggctgctg
      901 ccgctgcctc ctcagcctct ccagtctcca gggccatagg cagcaccgct aagccccagg
      961 agtctcccaa agggaagagg aagttggatt tgaaccaaga agaaaagaag acccccagca
     1021 aaccaccagc ccagctgtcg ccagctgttc ccaagagacc taaattgcca gctccaactc
     1081 gtaccccagc cacagcccca gtccctgccc gagcacaggg ggcagtgaca ggcaaacccc
     1141 gaggagaagg caccgagccc agacgacccc gagctggccc agaggagctg gggaagatcc
     1201 ttcagggtgt ggtagtggtg ctgagtggct tccagaaccc cttccgctcc gagctgcgag
     1261 ataaggccct agagcttggg gccaagtatc ggccagactg gacccgggac agcacgcacc
     1321 tcatctgtgc ctttgccaac acccccaagt acagccaggt cctaggcctg ggaggccgca
     1381 tcgtgcgtaa ggagtgggtg ctggactgtc accgcatgcg tcggcggctg ccctcccgga
     1441 ggtacctcat ggcagggcca ggttccagca gtgaggagga tgaggcctct cacagcggtg
     1501 gcagcggaga tgaagccccc aagcttcctc agaagcaacc ccagaccaaa accaagccca
     1561 ctcaggcagc tggacccagc tcaccccaga agcccccaac ccctgaagag accaaagcag
     1621 cctcaccagt gctccaggaa gatatagaca ttgagggggt acagtcagaa ggacaggaca
     1681 atggggcgga agattctggg gacacagagg atgagctgag gagggtggca gagcagaagg
     1741 aacacagact gccccctggc caggaggaga atggggaaga cccgtatgca ggctccacgg
     1801 atgagaacac ggacagtgag gaacaccagg agcctcctga tctgccagtc cctgagctcc
     1861 cagatttctt ccagggcaag cacttctttc tttacgggga gttccctggg gacgagcggc
     1921 ggaaactcat ccgatacgtc acagccttca atggggagct cgaggactat atgagtgacc
     1981 gggttcagtt tgtgatcaca gcacaggaat gggatcccag ctttgaggag gccctgatgg
     2041 acaacccctc cctggcattc gttcgtcccc gatggatcta cagttgcaat gagaagcaga
     2101 agttacttcc tcaccagctc tatggggtgg tgccgcaagc cgtcgagcac caccaccacc
     2161 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     2221 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     2281 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     2341 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2401 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2461 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2521 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2581 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2641 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2701 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     2761 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2821 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
     2881 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     2941 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     3001 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     3061 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     3121 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     3181 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     3241 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     3301 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     3361 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     3421 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     3481 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     3541 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     3601 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     3661 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     3721 tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3781 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3841 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3901 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     3961 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4021 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4081 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4141 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4201 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4261 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4321 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4381 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4441 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4501 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4561 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4621 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     4681 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     4741 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4801 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     4861 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     4921 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     4981 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     5041 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     5101 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     5161 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     5221 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     5281 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     5341 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     5401 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     5461 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     5521 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     5581 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     5641 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     5701 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     5761 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     5821 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5881 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     5941 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     6001 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     6061 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     6121 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     6181 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     6241 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     6301 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     6361 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     6421 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     6481 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     6541 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     6601 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     6661 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     6721 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     6781 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     6841 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     6901 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     6961 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     7021 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     7081 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaac
//
