LOCUS       sequence-404959-        9503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of PAR-deficient mXRCC1pd coupled to eGFP under
            the control of a EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9503)
  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 9503)
  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: pEF1a-mXRCC1pd-eGFP
FEATURES             Location/Qualifiers
     source          1..9503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(2047..2049,2050..2052,2053..2763)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2828..2949
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2839..2858)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2893..2912
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3064..3110
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3140..3276
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3321..3392
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3618..4206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3698..3717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4706..5915
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5752..5771
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5919..6518
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5919..5938)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6415..6435
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6545..6593
                     /note="synthetic polyadenylation signal"
     CDS             join(6859..6927,6928..7719)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7077..7096)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7732..7868
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8020..8091
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8096..8142)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8160..8208
                     /note="synthetic polyadenylation signal"
     promoter        8245..9423
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          8476..9414
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     9371..9391
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 gaattcagct agcagtgtac aatgccagaa atttcacttc ggcatgtagt ctcatgttct
       61 tcacaagatt ctacacactg cgctgagaac ctactgaaag cagataccta tagaaaatgg
      121 agagctgcga aagccggtga aaagacaatt tccgtagtgt tgcaactgga aaaggaagaa
      181 caaattcatt cagtcgatat cggaaacgac gggtccgcgt tcgtcgaagt actcgtcgga
      241 tcatcagccg gcggcgcaac tgccggagag caagattacg aagttctcct cgtgacgtct
      301 tcatttatgt cacctagtga atcaagatca ggtgccaacc caaatagagt acgtatcttc
      361 gggcccgata aacttgtgcg agccgctgca gaaaagcgat gggatagagt gaagatagtt
      421 tgttcccaac cctattctaa agattcccct tatggtctaa gcttcgttaa attccattct
      481 cccccagata aggacgaagc tgaagccact agtcaaaagg taaccgtcac taaactggga
      541 caattcagag tcaaggaaga agacgattcc gctaatgctc tcgcacccgg tgcattgttc
      601 tttgcacgga ttaataaaac cagcagtgct tccacctcag atcctgccgg tccgtcctac
      661 gccgccgcca cccttcaagc tgccgctgcc gcgagctctg cttcccccgt accaaaagtg
      721 gttgggtcca gcgcaaaacc ccaagaacca ccaaagggca aacgaaagct tgatctctca
      781 cttgaggatc gaaagccacc ttctaagcca tctgccggcc ctgccactct gaaacgtcct
      841 aagctgtccg tgccatcccg gacgcctgca gctgctcctg cttctacgcc cgcgcaacgc
      901 gcagtgcccg gtaaacctag aggtgaggga actgaacccc gaggcgccag gacaggaccg
      961 caggaactag ggaaaattct ccagggagtc gtagtcgtcc tgagcgggtt tcaaaatcct
     1021 tttcgatctg aactgagaga taaagctctc gagttgggcg ctaaataccg acccgattgg
     1081 acacctgatt ctacccacct gatatgcgct ttcgctaata cacctaagta ttcacaagtg
     1141 ttggggctgg gtggacgcat agttagaaag gaatgggttc tagattgcca tcatatgcgc
     1201 cggcgtcttc cttctaggag atatttgatg gctggactcg gttcatctag cgaagatgag
     1261 ggcgatagcc attccgaatc tggagaggat gaggcaccta aactgcctca gaaacgccca
     1321 caaccaaagg ctaaaactca agccgccggg ccatcttccc cacccagacc ccctactcct
     1381 aaggaaacaa aggcccccag ccccggccct caagataact ctgataccga aggtgaggaa
     1441 tctgagggta gagataatgg cgcggaagac agcggtgata cagaagacga gcttcgcaga
     1501 gtcgcaaagc aacgggagca gagacaacca cccgctcccg aagaaaacgg agaagatcct
     1561 tacgccggaa gcactgacga aaataccgat tcagaaactc cttccgaagc cgatctcccc
     1621 attcctgaac tgcctgattt ctttgaaggg aagcatttct ttctctacgg agaatttccg
     1681 ggagacgaac gtcgccggct gattaggtat gtcacggcat ttaacgggga actggaagat
     1741 tacatgaacg aacgagtaca atttgtgata actgcacaag aatgggatcc aaatttcgaa
     1801 gaagcgttga tggagaatcc gtcacttgca tttgttaggc caaggtggat ttattcttgc
     1861 aacgagaaac agaaactcct tcctcaccaa ctgtacggcg ttgttccaca agcggtggaa
     1921 cttaaactgc ggattctcca atccacagtc cccagagctc gagatccccc agtggctaca
     1981 gtagaactga aactcaggat ccttcaatcc acagtgccca gagctagaga tcccccagtt
     2041 gcaacaatgg tctcaaaagg ggaagaactt ttcacaggag tcgttccaat attggtagaa
     2101 ctcgatggtg atgttaatgg tcataaattt tcagtctctg gtgaagggga aggtgacgct
     2161 acatatggga aactcactct caaatttatt tgtactacag gaaaactccc tgtaccttgg
     2221 ccaactttgg ttactactct tacttatgga gtccaatgct tttctcgtta tcctgatcat
     2281 atgaaacaac atgatttctt taaatcagcg atgcctgagg ggtatgtgca agaaaggaca
     2341 atattcttca aagacgatgg taattataaa acaagggcag aagtaaaatt tgaaggagat
     2401 acgcttgtca atcgaataga attgaaggga attgatttta aagaagatgg caatatcctc
     2461 ggtcataaac tcgaatataa ttacaatagt cataatgtat acattatggc agataaacag
     2521 aagaatggca ttaaagtcaa ttttaaaatt cggcataata ttgaagacgg gtccgtccaa
     2581 cttgcagacc attatcaaca aaatacaccg attggagatg gtccagttct actccctgat
     2641 aatcattatc tatctacgca atctgctcta tccaaggatc caaatgagaa aagagaccat
     2701 atggtgctcc tcgaatttgt cacagcggcg ggcataaccc ttgggatgga tgaactctat
     2761 aaatagacga gatctaaggc tagcttgact gactgagata cagcgtacct tcagctcaca
     2821 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2881 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2941 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     3001 gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta
     3061 tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg
     3121 gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     3181 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     3241 ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga
     3301 tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca
     3361 gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa
     3421 acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac
     3481 gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt
     3541 acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3601 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3661 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3721 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3781 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3841 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3901 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3961 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4021 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4081 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4141 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4201 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4261 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4321 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4381 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4441 gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg
     4501 atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc
     4561 cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga
     4621 tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga
     4681 ggaatatcga aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc
     4741 gcccccctcg tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc
     4801 cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc
     4861 agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct
     4921 ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat
     4981 ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag
     5041 ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact
     5101 tggtgagtag cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg
     5161 gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc
     5221 ctgaactggg ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga
     5281 agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc
     5341 aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg
     5401 gctgggcacc atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg
     5461 tcgtctgttg cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg
     5521 ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt
     5581 ggggccacct gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg
     5641 gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag
     5701 tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc
     5761 ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt
     5821 ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagacttgt aaattgtccg
     5881 ctaaattctg gccgtttttg gcttttttgt tagacaacat gaccgagtac aagcccacgg
     5941 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     6001 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     6061 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     6121 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     6181 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     6241 agatggaggg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     6301 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     6361 tggaggccgc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     6421 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     6481 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctaggc tagtatgtaa gcctagtctt
     6541 agataataaa atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg
     6601 ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc
     6661 gttgcttggc agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga
     6721 tattgtactc aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag
     6781 tctgatcggt caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga
     6841 tcagcaggag gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6901 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6961 gaagatcagt tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7021 cttgagagtt ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta
     7081 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     7141 tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc
     7201 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     7261 ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     7321 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     7381 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc
     7441 gaactactta ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt
     7501 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7561 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7621 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7681 atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg
     7741 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     7801 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     7861 atccttaaga tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc
     7921 gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta
     7981 tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca
     8041 caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa
     8101 aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac
     8161 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg
     8221 atcgagtgta gccagatctc gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     8281 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
     8341 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     8401 gggggagaac cgtatataag tgcactagtc gccgtgaacg ttctttttcg caacgggttt
     8461 gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt
     8521 tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc
     8581 gagcttcggg ttggaagtgg gtgggagagt tcgtggcctt gcgcttaagg agccccttcg
     8641 cctcgtgctt gagttgtggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg
     8701 caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga
     8761 cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatcagcac
     8821 actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca
     8881 tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa
     8941 gctgcccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg
     9001 gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct
     9061 gctgcaggga gcacaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc
     9121 acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac
     9181 cgggcgccgt ccaggcacct cgattagttc tccagctttt ggagtacgtc gtctttaggt
     9241 tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt
     9301 aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct
     9361 tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg
     9421 tgaaaactac ccctaaaagc caaaagatct ttgtcgatcc taccatccac tcgacacacc
     9481 cgccagcggc cgctgccaag ctt
//
