LOCUS       sequence-209210-        7406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of BAP1-C91A (catalytically inactive mutant) in 
            mammalian cells, gene has an introduced siRNA-resistance against 
            siBAP1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7406)
  AUTHORS   Okonska A, Buhler S, Rao V, Ronner M, Blijlevens M, van der 
            Meulen-Muileman IH, de Menezes RX, Wipplinger M, Oehl K, Smit EF, 
            Weder W, Stahel RA, Penengo L, van Beusechem VW, Felley-Bosco E
  TITLE     Functional genomic screen in mesothelioma reveals that loss of 
            function of BRCA1-associated-protein-1 induces chemoresistance to 
            ribonucleotide reductase inhibition.
  JOURNAL   Mol Cancer Ther. 2019 Oct 16. pii: 1535-7163.MCT-19-0356. doi: 
            10.1158/1535-7163.MCT-19-0356.
  PUBMED    31619462
REFERENCE   2  (bases 1 to 7406)
  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: "Mol Cancer 
            Ther. 2019 Oct 16. pii: 1535-7163.MCT-19-0356. doi: 
            10.1158/1535-7163.MCT-19-0356."
COMMENT     Sequence Label: BAP1-C91A (siRNA resistant)
FEATURES             Location/Qualifiers
     source          1..7406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     589..613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          753..885
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     868..887
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        930..948
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2142..2153
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     polyA_signal    3224..3345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3235..3254)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3289..3308
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3531..3986
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3618..3637)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3828..3849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        4103..4294
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4126..4146)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4144..4441
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4453..5052
                     /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(4453..4472)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4949..4969
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5118..5166
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5207..5226)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5326..5348
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5386..5404)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5472..5576
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5577..5645,5646..6437)
                     /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(5795..5814)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6608..7196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7097..7116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      601 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      661 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agtgactctc ttaaggtagc
      721 cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
      781 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      841 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
      901 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct agcatgaata
      961 agggctggct ggagctggag agcgacccag gcctcttcac cctgctcgtg gaagatttcg
     1021 gtgtcaaggg ggtgcaagtg gaggagatct acgaccttca gagcaaatgt cagggccctg
     1081 tatatggatt tatcttcctg ttcaaatgga tcgaagagcg ccggtcccgg cgaaaggtct
     1141 ctaccttggt ggatgatacg tccgtgattg atgatgatat tgtgaataac atgttctttg
     1201 cccaccagct gatacccaac tctgctgcaa ctcatgcctt gctgagcgtg ctcctgaact
     1261 gcagcagcgt ggacctggga cccaccctga gtcgcatgaa ggacttcacc aagggtttca
     1321 gccctgagag caaaggatat gcgattggca atgccccgga gttggccaag gcccataata
     1381 gccatgccag gcccgagcca cgccacctcc ctgagaagca gaatggcctt agtgcagtgc
     1441 ggaccatgga ggcgttccac tttgtcagct atgtgcctat cacaggccgg ctctttgagc
     1501 tggatgggct gaaggtctac cccattgacc atgggccctg gggggaggac gaggagtgga
     1561 cagacaaggc ccggcgggtc atcatggagc gtatcggcct cgccactgca ggggagccct
     1621 accacgacat ccgcttcaac ctgatggcag tggtgcccga ccgcaggatc aagtatgagg
     1681 ccaggctgca tgtgctgaag gtgaaccgtc agacagtact agaggctctg cagcagctga
     1741 taagagtaac acagccagag ctgattcaga cccacaagtc tcaagagtca cagctgcctg
     1801 aggagtccaa gtcagccagc aacaagtccc cgctggtgct ggaagcaaac agggcccctg
     1861 cagcctctga gggcaaccac acagatggtg cagaggaggc ggctggttca tgcgcacaag
     1921 ccccatccca cagccctccc aacaaaccca agctagtggt gaagcctcca ggcagcagcc
     1981 tcaatggggt tcaccccaac cccactccca ttgtccagcg gctgccggcc tttctagaca
     2041 atcacaatta tgccaagtcc cccatgcagg aggaagaaga cctggcggca ggtgtgggcc
     2101 gcagccgagt tccagtccgc ccaccccagc agtactcaga tgatgaggat gactatgagg
     2161 atgacgagga ggatgacgtg cagaacacca actctgccct taggtataag gggaagggaa
     2221 cagggaagcc aggggcattg agcggttctg ctgatgggca actgtcagtg ctgcagccca
     2281 acaccatcaa cgtcttggct gagaagctca aagagtccca gaaggacctc tcaattcctc
     2341 tgtccatcaa gactagcagc ggggctggga gtccggctgt ggcagtgccc acacactcgc
     2401 agccctcacc cacccccagc aatgagagta cagacacggc ctctgagatc ggcagtgctt
     2461 tcaactcgcc actgcgctcg cctatccgct cagccaaccc gacgcggccc tccagccctg
     2521 tcacctccca catctccaag gtgctttttg gagaggatga cagcctgctg cgtgttgact
     2581 gcatacgcta caatcgagcc gtgcgagatc tgggtcctgt catcagcaca ggcctgctgc
     2641 acctggctga ggatggggtg ctgagtcccc tggcgctgac agagggtggg aagggttcct
     2701 cgccctccat cagaccaatc caaggcagcc aggggtccag cagcccagtg gagaaggagg
     2761 tcgtggaagc cacggacagc agagagaaga cggggatggt gaggcctggc gagcccttga
     2821 gtggggagaa atactcaccc aaggagctgc tggcactgct gaagtgtgtg gaggctgaga
     2881 ttgcaaacta tgaggcgtgc ctcaaggagg aggtagagaa gaggaagaag ttcaagattg
     2941 atgaccagag aaggacccac aactacgatg agttcatctg cacctttatc tccatgctgg
     3001 ctcaggaagg catgctggcc aacctagtgg agcagaacat ctccgtgcgg cggcgccaag
     3061 gggtcagcat cggccggctc cacaagcagc ggaagcctga ccggcggaaa cgctctcgcc
     3121 cctacaaggc caagcgccag tgagaattca cgcgtggtac ctctagagtc gacccgggcg
     3181 gccgcttccc tttagtgagg gttaatgctt cgagcagaca tgataagata cattgatgag
     3241 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3301 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3361 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     3421 ctctacaaat gtggtaaaat ccgataagga tcgatccggg ctggcgtaat agcgaagagg
     3481 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     3541 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3601 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3661 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3721 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3781 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3841 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3901 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3961 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     4021 ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata acctctgaaa
     4081 gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4141 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4201 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4261 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4321 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4381 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4441 gaggagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     4501 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     4561 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     4621 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     4681 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     4741 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     4801 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     4861 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     4921 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     4981 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     5041 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     5101 catgcatcga tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     5161 tgtgtgaatc gatagcgata aggatccgcg tatggtgcac tctcagtaca atctgctctg
     5221 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5281 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5341 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5401 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5461 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5521 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5581 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5641 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5701 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5761 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5821 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5881 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5941 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6001 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6061 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6121 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6181 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6241 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6301 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6361 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6421 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6481 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6541 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6601 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6661 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6721 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6781 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6841 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6901 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6961 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7021 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7081 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7141 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7201 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatggctc
     7261 gacagatctt caatattggc cattagccat attattcatt ggttatatag cataaatcaa
     7321 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     7381 ctcatgtcca atatgaccgc catgtt
//
