LOCUS       sequence-330065-        9721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding for 2 gRNAs targeting the human BAX gene and a CMV 
            driven nuclease Cas9 followed by self-cleaving EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9721)
  AUTHORS   Tigano M, Vargas DC, Tremblay-Belzile S, Fu Y, Sfeir A
  TITLE     Nuclear sensing of breaks in mitochondrial DNA enhances immune 
            surveillance.
  JOURNAL   Nature. 2021 Mar;591(7850):477-481. doi: 10.1038/s41586-021-03269-w.
            Epub 2021 Feb 24.
  PUBMED    33627873
REFERENCE   2  (bases 1 to 9721)
  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/s41586-021-03269-w"; journalName: "Nature"; date: 
            "2021-03"; volume: "591"; issue: "7850"; pages: "477-481"
COMMENT     Sequence Label: pSpCas9nucl-T2A-EGFP - BAXgRNA1- BAXgRNA2
FEATURES             Location/Qualifiers
     source          1..9721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        437..685
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        707..782
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        873..1158
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1160..1437
                     /label=chicken beta-actin promoter
     intron          1438..1665
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1677..1686
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1686..1751
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1758..1778
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1803..5903
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1803..5903
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5904..5951
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5967..6020
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(6021..6023,6024..6734)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6021..6023,6024..6734)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6324..6343)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6671..6692
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6762..6779)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6768..6975
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6984..7124
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6984..7113
                     /label=AAV2 ITR
     rep_origin      7199..7654
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7286..7305)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7496..7517
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7671..7690)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7790..7812
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7850..7868)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7936..8040
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8041..8109,8110..8901)
                     /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(8259..8278)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9072..9660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9561..9580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ctgcaggatg attgccgccg gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      421 gacaaatggc tctagagagg gcctatttcc catgattcct tcatatttgc atatacgata
      481 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
      541 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
      601 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
      661 tatcttgtgg aaaggacgaa acaccgtctg acggcaactt caactggttt tagagctaga
      721 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
      781 gcttttttgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg tttttagcgc
      841 gtgcgccaat tctgcagaca aatggcggta cccgttacat aacttacggt aaatggcccg
      901 cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact
      961 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1021 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     1081 cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     1141 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     1201 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     1261 ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc
     1321 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
     1381 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
     1441 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
     1501 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
     1561 ggctgtaatt agctgagcaa gaggtaaggg tttaagggat ggttggttgg tggggtatta
     1621 atgtttaatt acctggagca cctgcctgaa atcacttttt ttcaggttgg accggtgcca
     1681 ccatggacta taaggaccac gacggagact acaaggatca tgatattgat tacaaagacg
     1741 atgacgataa gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag
     1801 ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
     1861 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     1921 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
     1981 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     2041 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     2101 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     2161 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     2221 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     2281 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     2341 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     2401 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     2461 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     2521 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     2581 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     2641 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     2701 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     2761 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     2821 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     2881 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     2941 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     3001 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     3061 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     3121 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     3181 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     3241 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     3301 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     3361 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     3421 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     3481 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     3541 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     3601 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     3661 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     3721 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     3781 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     3841 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     3901 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     3961 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     4021 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     4081 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     4141 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     4201 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     4261 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata
     4321 tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg
     4381 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     4441 actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga
     4501 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     4561 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     4621 ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca
     4681 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     4741 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     4801 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     4861 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     4921 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     4981 ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg
     5041 ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     5101 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg
     5161 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
     5221 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     5281 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     5341 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     5401 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
     5461 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     5521 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     5581 agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     5641 tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc
     5701 ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
     5761 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
     5821 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
     5881 acctgtctca gctgggaggc gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa
     5941 aaaagaaaaa ggaattcggc agtggagagg gcagaggaag tctgctaaca tgcggtgacg
     6001 tcgaggagaa tcctggccca gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     6061 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     6121 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     6181 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     6241 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     6301 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     6361 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     6421 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     6481 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     6541 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     6601 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     6661 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     6721 acgagctgta caaggaattc taactagagc tcgctgatca gcctcgactg tgccttctag
     6781 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     6841 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     6901 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag
     6961 caggcatgct ggggagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     7021 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     7081 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     7141 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     7201 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     7261 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     7321 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     7381 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     7441 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     7501 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     7561 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     7621 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     7681 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     7741 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     7801 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     7861 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     7921 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     7981 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     8041 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     8101 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     8161 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     8221 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     8281 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     8341 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     8401 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     8461 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     8521 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     8581 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     8641 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     8701 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc
     8761 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     8821 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     8881 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     8941 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     9001 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     9061 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     9121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     9181 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     9241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     9301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     9361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     9421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     9481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     9541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     9601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     9661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     9721 t
//
