LOCUS       sequence-421625-        7780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV driven PARP1-E988K -eGFP expressing construct with IRES-Neomycin
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7780)
  AUTHORS   Krastev DB, Li S, Sun Y, Wicks AJ, Hoslett G, Weekes D, Badder LM, 
            Knight EG, Marlow R, Pardo MC, Yu L, Talele TT, Bartek J, Choudhary 
            JS, Pommier Y, Pettitt SJ, Tutt ANJ, Ramadan K, Lord CJ
  TITLE     The ubiquitin-dependent ATPase p97 removes cytotoxic trapped PARP1 
            from chromatin.
  JOURNAL   Nat Cell Biol. 2022 Jan;24(1):62-73. doi: 
            10.1038/s41556-021-00807-6. Epub 2022 Jan 10.
  PUBMED    35013556
REFERENCE   2  (bases 1 to 7780)
  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"; doi: 
            "10.1038/s41556-021-00807-6"; journalName: "Nat Cell Biol"; date: 
            "2022-01"; volume: "24"; issue: "1"; pages: "62-73"
COMMENT     Sequence Label: pEGFP-N1-PARP1-E988K
FEATURES             Location/Qualifiers
     source          1..7780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             606..629
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3720..3729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3726..3728,3729..3731,3732..4445)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3726..3728,3729..3731,3732..4442)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3771..3792)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4032..4051)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4379..4400
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4568..4689
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4605..4624)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4659..4678
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4696..5151)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4833..4854)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5045..5064
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5178..5282
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5284..5641
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5284..5475
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5307..5327)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5492..5627
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5676..6470
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5676..6470
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5730..5749)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6340..6359
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6658..6677)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6702..6749
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7078..7666
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7567..7586
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      601 ccatggacta caaagacgat gacgacaagc ttgccgccgc gaattcaatg gcggagtctt
      661 cggataagct ctatcgagtc gagtacgcca agagcgggcg cgcctcttgc aagaaatgca
      721 gcgagagcat ccccaaggac tcgctccgga tggccatcat ggtgcagtcg cccatgtttg
      781 atggaaaagt cccacactgg taccacttct cctgcttctg gaaggtgggc cactccatcc
      841 ggcaccctga cgttgaggtg gatgggttct ctgagcttcg gtgggatgac cagcagaaag
      901 tcaagaagac agcggaagct ggaggagtga caggcaaagg ccaggatgga attggtagca
      961 aggcagagaa gactctgggt gactttgcag cagagtatgc caagtccaac agaagtacgt
     1021 gcaaggggtg tatggagaag atagaaaagg gccaggtgcg cctgtccaag aagatggtgg
     1081 acccggagaa gccacagcta ggcatgattg accgctggta ccatccaggc tgctttgtca
     1141 agaacaggga ggagctgggt ttccggcccg agtacagtgc gagtcagctc aagggcttca
     1201 gcctccttgc tacagaggat aaagaagccc tgaagaagca gctcccagga gtcaagagtg
     1261 aaggaaagag aaaaggcgat gaggtggatg gagtggatga agtggcgaag aagaaatcta
     1321 aaaaagaaaa agacaaggat agtaagcttg aaaaagccct aaaggctcag aacgacctga
     1381 tctggaacat caaggacgag ctaaagaaag tgtgttcaac taatgacctg aaggagctac
     1441 tcatcttcaa caagcagcaa gtgccttctg gggagtcggc gatcttggac cgagtagctg
     1501 atggcatggt gttcggtgcc ctccttccct gcgaggaatg ctcgggtcag ctggtcttca
     1561 agagcgatgc ctattactgc actggggacg tcactgcctg gaccaagtgt atggtcaaga
     1621 cacagacacc caaccggaag gagtgggtaa ccccaaagga attccgagaa atctcttacc
     1681 tcaagaaatt gaaggttaaa aagcaggacc gtatattccc cccagaaacc agcgcctccg
     1741 tggcggccac gcctccgccc tccacagcct cggctcctgc tgctgtgaac tcctctgctt
     1801 cagcagataa gccattatcc aacatgaaga tcctgactct cgggaagctg tcccggaaca
     1861 aggatgaagt gaaggccatg attgagaaac tcggggggaa gttgacgggg acggccaaca
     1921 aggcttccct gtgcatcagc accaaaaagg aggtggaaaa gatgaataag aagatggagg
     1981 aagtaaagga agccaacatc cgagttgtgt ctgaggactt cctccaggac gtctccgcct
     2041 ccaccaagag ccttcaggag ttgttcttag cgcacatctt gtccccttgg ggggcagagg
     2101 tgaaggcaga gcctgttgaa gttgtggccc caagagggaa gtcaggggct gcgctctcca
     2161 aaaaaagcaa gggccaggtc aaggaggaag gtatcaacaa atctgaaaag agaatgaaat
     2221 taactcttaa aggaggagca gctgtggatc ctgattctgg actggaacac tctgcgcatg
     2281 tcctggagaa aggtgggaag gtcttcagtg ccacccttgg cctggtggac atcgttaaag
     2341 gaaccaactc ctactacaag ctgcagcttc tggaggacga caaggaaaac aggtattgga
     2401 tattcaggtc ctggggccgt gtgggtacgg tgatcggtag caacaaactg gaacagatgc
     2461 cgtccaagga ggatgccatt gagcacttca tgaaattata tgaagaaaaa accgggaacg
     2521 cttggcactc caaaaatttc acgaagtatc ccaaaaagtt ctaccccctg gagattgact
     2581 atggccagga tgaagaggca gtgaagaagc tgacagtaaa tcctggcacc aagtccaagc
     2641 tccccaagcc agttcaggac ctcatcaaga tgatctttga tgtggaaagt atgaagaaag
     2701 ccatggtgga gtatgagatc gaccttcaga agatgccctt ggggaagctg agcaaaaggc
     2761 agatccaggc cgcatactcc atcctcagtg aggtccagca ggcggtgtct cagggcagca
     2821 gcgactctca gatcctggat ctctcaaatc gcttttacac cctgatcccc cacgactttg
     2881 ggatgaagaa gcctccgctc ctgaacaatg cagacagtgt gcaggccaag gtggaaatgc
     2941 ttgacaacct gctggacatc gaggtggcct acagtctgct caggggaggg tctgatgata
     3001 gcagcaagga tcccatcgat gtcaactatg agaagctcaa aactgacatt aaggtggttg
     3061 acagagattc tgaagaagcc gagatcatca ggaagtatgt taagaacact catgcaacca
     3121 cacacaatgc gtatgacttg gaagtcatcg atatctttaa gatagagcgt gaaggcgaat
     3181 gccagcgtta caagcccttt aagcagcttc ataaccgaag attgctgtgg cacgggtcca
     3241 ggaccaccaa ctttgctggg atcctgtccc agggtcttcg gatagccccg cctgaagcgc
     3301 ccgtgacagg ctacatgttt ggtaaaggga tctatttcgc tgacatggtc tccaagagtg
     3361 ccaactactg ccatacgtct cagggagacc caataggctt aatcctgttg ggagaagttg
     3421 cccttggaaa catgtatgaa ctgaagcacg cttcacatat cagcaagtta cccaagggca
     3481 agcacagtgt caaaggtttg ggcaaaacta cccctgatcc ttcagctaac attagtctgg
     3541 atggtgtaga cgttcctctt gggaccggga tttcatctgg tgtgaatgac acctctctac
     3601 tatataacaa gtacattgtc tatgatattg ctcaggtaaa tctgaagtat ctgctgaaac
     3661 tgaaattcaa ttttaagacc tccctgtggt cggtaccgcg ggcccgggat ccaccggtcg
     3721 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     3781 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     3841 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3901 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     3961 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     4021 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     4081 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     4141 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     4201 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     4261 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     4321 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     4381 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     4441 agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     4501 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4561 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4621 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4681 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4741 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4801 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4861 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4921 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4981 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     5041 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     5101 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     5161 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5221 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5281 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     5341 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     5401 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5461 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5521 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5581 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5641 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5701 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5761 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5821 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     5881 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5941 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     6001 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     6061 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     6121 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     6181 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     6241 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     6301 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     6361 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6421 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6481 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6541 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6601 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     6661 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     6721 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     6781 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     6841 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     6901 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     6961 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7021 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7081 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7141 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7201 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7261 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7321 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7381 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7441 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7501 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7561 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7621 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7681 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7741 atcccctgat tctgtggata accgtattac cgccatgcat
//
