LOCUS       sequence-298080-        7424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HA-tagged p.C91S Mutant BAP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7424)
  AUTHORS   Niersch J, Vega-Rubin-de-Celis S, Bazarna A, Mergener S, Jendrossek 
            V, Siveke JT, Pena-Llopis S
  TITLE     A BAP1 synonymous mutation results in exon skipping, loss of 
            function and worse patient prognosis.
  JOURNAL   iScience. 2021 Feb 12;24(3):102173. doi: 10.1016/j.isci.2021.102173.
            eCollection 2021 Mar 19.
  PUBMED    33681728
REFERENCE   2  (bases 1 to 7424)
  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: "iScience.";
            date: "2021-02-12"; volume: "24(3):102173. doi"; pages: " 
            10.1016/j.isci.2021.102173. eCollection 2021 Mar 19"
COMMENT     Sequence Label: pBABE-hygro-BAP1-C91S-HA
FEATURES             Location/Qualifiers
     source          1..7424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    2556..2567
                     /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)"
     CDS             3555..3581
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3614..3634)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3619..3948
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3799..3934
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4022..5047
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4022..5047
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     5564..5583
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             5845..6314
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(join(6429..7220,7221..7289))
                     /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     7052..7071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7290..7394)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc cgccaccatg aataagggct
     1381 ggctggagct ggagagcgac ccaggcctct tcaccctgct cgtggaagat ttcggtgtca
     1441 agggggtgca agtggaggag atctacgacc ttcagagcaa atgtcagggc cctgtgtatg
     1501 gatttatctt cctgttcaaa tggatcgaag agcgccggtc ccggcgaaag gtctctacct
     1561 tggtggatga tacgtccgtg attgatgatg atattgtgaa taacatgttc tttgcccacc
     1621 agctgatacc caactcgagt gcaactcatg ccttgctgag cgtgctcctg aactgcagca
     1681 gcgtggacct gggacccacc ctgagtcgca tgaaggactt caccaagggt ttcagccctg
     1741 agagcaaagg atatgcgatt ggcaatgccc cggagttggc caaggcccat aatagccatg
     1801 ccaggcccga gccacgccac ctccctgaga agcagaatgg ccttagtgca gtgcggacca
     1861 tggaggcgtt ccactttgtc agctatgtgc ctatcacagg ccggctcttt gagctggatg
     1921 ggctgaaggt ctaccccatt gaccatgggc cctgggggga ggacgaggag tggacagaca
     1981 aggcccggcg ggtcatcatg gagcgtatcg gcctcgccac tgcaggggag ccctaccacg
     2041 acatccgctt caacctgatg gcagtggtgc ccgaccgcag gatcaagtat gaggccaggc
     2101 tgcatgtgct gaaggtgaac cgtcagacag tactagaggc tctgcagcag ctgataagag
     2161 taacacagcc agagctgatt cagacccaca agtctcaaga gtcacagctg cctgaggagt
     2221 ccaagtcagc cagcaacaag tccccgctgg tgctggaagc aaacagggcc cctgcagcct
     2281 ctgagggcaa ccacacagat ggtgcagagg aggcggctgg ttcatgcgca caagccccat
     2341 cccacagccc tcccaacaaa cccaagctag tggtgaagcc tccaggcagc agcctcaatg
     2401 gggttcaccc caaccccact cccattgtcc agcggctgcc ggcctttcta gacaatcaca
     2461 attatgccaa gtcccccatg caggaggaag aagacctggc ggcaggtgtg ggccgcagcc
     2521 gagttccagt ccgcccaccc cagcagtact cagatgatga ggatgactat gaggatgacg
     2581 aggaggatga cgtgcagaac accaactctg cccttaggta taaggggaag ggaacaggga
     2641 agccaggggc attgagcggt tctgctgatg ggcaactgtc agtgctgcag cccaacacca
     2701 tcaacgtctt ggctgagaag ctcaaagagt cccagaagga cctctcaatt cctctgtcca
     2761 tcaagactag cagcggggct gggagtccgg ctgtggcagt gcccacacac tcgcagccct
     2821 cacccacccc cagcaatgag agtacagaca cggcctctga gatcggcagt gctttcaact
     2881 cgccactgcg ctcgcctatc cgctcagcca acccgacgcg gccctccagc cctgtcacct
     2941 cccacatctc caaggtgctt tttggagagg atgacagcct gctgcgtgtt gactgcatac
     3001 gctacaaccg tgctgtccgt gatctgggtc ctgtcatcag cacaggcctg ctgcacctgg
     3061 ctgaggatgg ggtgctgagt cccctggcgc tgacagaggg tgggaagggt tcctcgccct
     3121 ccatcagacc aatccaaggc agccaggggt ccagcagccc agtggagaag gaggtcgtgg
     3181 aagccacgga cagcagagag aagacgggga tggtgaggcc tggcgagccc ttgagtgggg
     3241 agaaatactc acccaaggag ctgctggcac tgctgaagtg tgtggaggct gagattgcaa
     3301 actatgaggc gtgcctcaag gaggaggtag agaagaggaa gaagttcaag attgatgacc
     3361 agagaaggac ccacaactac gatgaattta tatgtacgtt catatcgatg ctggctcagg
     3421 aaggcatgct ggccaaccta gtggagcaga acatctccgt gcggcggcgc caaggggtca
     3481 gcatcggccg gctccacaag cagcggaagc ctgaccggcg gaaacgctct cgcccctaca
     3541 aggccaagcg ccagtaccca tacgatgttc cagattacgc ttgagaattc gccagcacag
     3601 tggtcgaccc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3661 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3721 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3781 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3841 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3901 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgccagc
     3961 tggggcgccc tctggtaagg ttgggaagcc ctgcagatct gatcccgggg ggcaatgaga
     4021 tatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     4081 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     4141 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     4201 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
     4261 tggggaattt agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     4321 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga
     4381 tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     4441 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     4501 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     4561 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     4621 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     4681 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     4741 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     4801 gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     4861 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     4921 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg
     4981 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     5041 ggaatagagt agatgccgac cgggatctat cgatgcggcg ggtgtggtgg ttacgcgtaa
     5101 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5161 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5221 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5281 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5341 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5401 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5461 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5521 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5581 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5641 caggggggcg gagcctatgg aaaaacgcca gcaacgcatc gataaaataa aagattttat
     5701 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     5761 aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat
     5821 caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     5881 gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga
     5941 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     6001 ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     6061 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     6121 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc
     6181 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     6241 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     6301 gcgggggtct ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt
     6361 tacattaaat ggccatagtg cggccgcaaa tcaatctaaa gtatatatga gtaaacttgg
     6421 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6481 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6541 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6601 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6661 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6721 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6781 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6841 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6901 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6961 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7021 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7081 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7141 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7201 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7261 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7321 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7381 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg cagc
//
