LOCUS       sequence-421624-        7777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV driven PARP1-delM43;F44I -eGFP expressing construct with 
            IRES-Neomycin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7777)
  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 7777)
  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-delM43;F44I
FEATURES             Location/Qualifiers
     source          1..7777
                     /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      3717..3726
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3723..3725,3726..3728,3729..4442)
                     /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(3723..3725,3726..3728,3729..4439)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3768..3789)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4029..4048)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4376..4397
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4565..4686
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4602..4621)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4656..4675
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4693..5148)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4830..4851)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5042..5061
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5175..5279
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5281..5638
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5281..5472
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5304..5324)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5489..5624
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5673..6467
                     /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             5673..6467
                     /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(5727..5746)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6337..6356
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6655..6674)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6699..6746
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7075..7663
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7564..7583
                     /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 cccattgatg
      781 gaaaagtccc acactggtac cacttctcct gcttctggaa ggtgggccac tccatccggc
      841 accctgacgt tgaggtggat gggttctctg agcttcggtg ggatgaccag cagaaagtca
      901 agaagacagc ggaagctgga ggagtgacag gcaaaggcca ggatggaatt ggtagcaagg
      961 cagagaagac tctgggtgac tttgcagcag agtatgccaa gtccaacaga agtacgtgca
     1021 aggggtgtat ggagaagata gaaaagggcc aggtgcgcct gtccaagaag atggtggacc
     1081 cggagaagcc acagctaggc atgattgacc gctggtacca tccaggctgc tttgtcaaga
     1141 acagggagga gctgggtttc cggcccgagt acagtgcgag tcagctcaag ggcttcagcc
     1201 tccttgctac agaggataaa gaagccctga agaagcagct cccaggagtc aagagtgaag
     1261 gaaagagaaa aggcgatgag gtggatggag tggatgaagt ggcgaagaag aaatctaaaa
     1321 aagaaaaaga caaggatagt aagcttgaaa aagccctaaa ggctcagaac gacctgatct
     1381 ggaacatcaa ggacgagcta aagaaagtgt gttcaactaa tgacctgaag gagctactca
     1441 tcttcaacaa gcagcaagtg ccttctgggg agtcggcgat cttggaccga gtagctgatg
     1501 gcatggtgtt cggtgccctc cttccctgcg aggaatgctc gggtcagctg gtcttcaaga
     1561 gcgatgccta ttactgcact ggggacgtca ctgcctggac caagtgtatg gtcaagacac
     1621 agacacccaa ccggaaggag tgggtaaccc caaaggaatt ccgagaaatc tcttacctca
     1681 agaaattgaa ggttaaaaag caggaccgta tattcccccc agaaaccagc gcctccgtgg
     1741 cggccacgcc tccgccctcc acagcctcgg ctcctgctgc tgtgaactcc tctgcttcag
     1801 cagataagcc attatccaac atgaagatcc tgactctcgg gaagctgtcc cggaacaagg
     1861 atgaagtgaa ggccatgatt gagaaactcg gggggaagtt gacggggacg gccaacaagg
     1921 cttccctgtg catcagcacc aaaaaggagg tggaaaagat gaataagaag atggaggaag
     1981 taaaggaagc caacatccga gttgtgtctg aggacttcct ccaggacgtc tccgcctcca
     2041 ccaagagcct tcaggagttg ttcttagcgc acatcttgtc cccttggggg gcagaggtga
     2101 aggcagagcc tgttgaagtt gtggccccaa gagggaagtc aggggctgcg ctctccaaaa
     2161 aaagcaaggg ccaggtcaag gaggaaggta tcaacaaatc tgaaaagaga atgaaattaa
     2221 ctcttaaagg aggagcagct gtggatcctg attctggact ggaacactct gcgcatgtcc
     2281 tggagaaagg tgggaaggtc ttcagtgcca cccttggcct ggtggacatc gttaaaggaa
     2341 ccaactccta ctacaagctg cagcttctgg aggacgacaa ggaaaacagg tattggatat
     2401 tcaggtcctg gggccgtgtg ggtacggtga tcggtagcaa caaactggaa cagatgccgt
     2461 ccaaggagga tgccattgag cacttcatga aattatatga agaaaaaacc gggaacgctt
     2521 ggcactccaa aaatttcacg aagtatccca aaaagttcta ccccctggag attgactatg
     2581 gccaggatga agaggcagtg aagaagctga cagtaaatcc tggcaccaag tccaagctcc
     2641 ccaagccagt tcaggacctc atcaagatga tctttgatgt ggaaagtatg aagaaagcca
     2701 tggtggagta tgagatcgac cttcagaaga tgcccttggg gaagctgagc aaaaggcaga
     2761 tccaggccgc atactccatc ctcagtgagg tccagcaggc ggtgtctcag ggcagcagcg
     2821 actctcagat cctggatctc tcaaatcgct tttacaccct gatcccccac gactttggga
     2881 tgaagaagcc tccgctcctg aacaatgcag acagtgtgca ggccaaggtg gaaatgcttg
     2941 acaacctgct ggacatcgag gtggcctaca gtctgctcag gggagggtct gatgatagca
     3001 gcaaggatcc catcgatgtc aactatgaga agctcaaaac tgacattaag gtggttgaca
     3061 gagattctga agaagccgag atcatcagga agtatgttaa gaacactcat gcaaccacac
     3121 acaatgcgta tgacttggaa gtcatcgata tctttaagat agagcgtgaa ggcgaatgcc
     3181 agcgttacaa gccctttaag cagcttcata accgaagatt gctgtggcac gggtccagga
     3241 ccaccaactt tgctgggatc ctgtcccagg gtcttcggat agccccgcct gaagcgcccg
     3301 tgacaggcta catgtttggt aaagggatct atttcgctga catggtctcc aagagtgcca
     3361 actactgcca tacgtctcag ggagacccaa taggcttaat cctgttggga gaagttgccc
     3421 ttggaaacat gtatgaactg aagcacgctt cacatatcag caagttaccc aagggcaagc
     3481 acagtgtcaa aggtttgggc aaaactaccc ctgatccttc agctaacatt agtctggatg
     3541 gtgtagacgt tcctcttggg accgggattt catctggtgt gaatgacacc tctctactat
     3601 ataacgagta cattgtctat gatattgctc aggtaaatct gaagtatctg ctgaaactga
     3661 aattcaattt taagacctcc ctgtggtcgg taccgcgggc ccgggatcca ccggtcgcca
     3721 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     3781 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     3841 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     3901 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     3961 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     4021 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     4081 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     4141 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     4201 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     4261 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     4321 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     4381 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     4441 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     4501 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     4561 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4621 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4681 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4741 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4801 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4861 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4921 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4981 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5041 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     5101 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     5161 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5221 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     5281 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     5341 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     5401 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5461 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5521 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5581 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     5641 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5701 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5761 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5821 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5881 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5941 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     6001 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     6061 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6121 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6181 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6241 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6301 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6361 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6421 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6481 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6541 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6601 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     6661 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     6721 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     6781 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6841 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6901 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     6961 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7021 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7081 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7141 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7201 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7261 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7321 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7381 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7441 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7501 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7561 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7621 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7681 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7741 ccctgattct gtggataacc gtattaccgc catgcat
//
