LOCUS       sequence-254648-       10359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG promoter expression plasmid for 
            NLS-hSpCas9n(D10A)-NLS-SH3-3xFLAG-NLS-UGI-P2A-EGFP (Target-AID 
            without pmCDA1). .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10359)
  AUTHORS   Grunewald J, Zhou R, Iyer S, Lareau CA, Garcia SP, Aryee MJ, Joung 
            JK
  TITLE     CRISPR DNA base editors with reduced RNA off-target and self-editing
            activities.
  JOURNAL   Nat Biotechnol. 2019 Sep 2. pii: 10.1038/s41587-019-0236-6. doi: 
            10.1038/s41587-019-0236-6.
  PUBMED    31477922
REFERENCE   2  (bases 1 to 10359)
  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: "Nat 
            Biotechnol. 2019 Sep 2. pii: 10.1038/s41587-019-0236-6. doi: 
            10.1038/s41587-019-0236-6."
COMMENT     Sequence Label: nCas9-UGI control for Target-AID (pRZ1047)
FEATURES             Location/Qualifiers
     source          1..10359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          135..1151
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1075..1097
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1159..1178
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1244..1253
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1256..1276
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1301..1327,1328..1330,1331..5404)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1301..5404
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5417..5437
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5645..5710
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5720..5740
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5747..5998
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6017..6073
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(6074..6076,6077..6079,6080..6793)
                     /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(6074..6076,6077..6079,6080..6790)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6119..6140)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6380..6399)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6727..6748
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6880..6899)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6945..7000
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6999..7018)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7361..7377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7361..7377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7374..7396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7385..7401
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7409..7415,7416..7433,7434..7439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7454..7475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7534..7729
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7580..7715
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7735..7869
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7772..7791)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7826..7845
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7937..7954)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8108..8696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8188..8207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8867..9658,9659..9727))
                     /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     9490..9509
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9728..9832)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        9863..10242
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggcnnnnn gmggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg
       61 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
      121 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg
      181 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
      241 acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc
      301 tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct
      361 cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg
      421 gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg
      481 ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct
      541 gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc
      601 aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc
      661 tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg
      721 tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc
      781 cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat
      841 cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga
      901 ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg
      961 aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc
     1021 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
     1081 gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct
     1141 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
     1201 aattctgcag tcgacggtac cgcgggcccg ggatccaccg gtcgccacca tggcaccgaa
     1261 gaagaagcgt aaagtcggaa tccacggagt tcctgcggca atggacaaga agtactccat
     1321 tgggctcgct atcggcacaa acagcgtcgg ttgggccgtc attacggacg agtacaaggt
     1381 gccgagcaaa aaattcaaag ttctgggcaa taccgatcgc cacagcataa agaagaacct
     1441 cattggcgcc ctcctgttcg actccgggga gacggccgaa gccacgcggc tcaaaagaac
     1501 agcacggcgc agatataccc gcagaaagaa tcggatctgc tacctgcagg agatctttag
     1561 taatgagatg gctaaggtgg atgactcttt cttccatagg ctggaggagt cctttttggt
     1621 ggaggaggat aaaaagcacg agcgccaccc aatctttggc aatatcgtgg acgaggtggc
     1681 gtaccatgaa aagtacccaa ccatatatca tctgaggaag aagcttgtag acagtactga
     1741 taaggctgac ttgcggttga tctatctcgc gctggcgcat atgatcaaat ttcggggaca
     1801 cttcctcatc gagggggacc tgaacccaga caacagcgat gtcgacaaac tctttatcca
     1861 actggttcag acttacaatc agcttttcga agagaacccg atcaacgcat ccggagttga
     1921 cgccaaagca atcctgagcg ctaggctgtc caaatcccgg cggctcgaaa acctcatcgc
     1981 acagctccct ggggagaaga agaacggcct gtttggtaat cttatcgccc tgtcactcgg
     2041 gctgaccccc aactttaaat ctaacttcga cctggccgaa gatgccaagc ttcaactgag
     2101 caaagacacc tacgatgatg atctcgacaa tctgctggcc cagatcggcg accagtacgc
     2161 agaccttttt ttggcggcaa agaacctgtc agacgccatt ctgctgagtg atattctgcg
     2221 agtgaacacg gagatcacca aagctccgct gagcgctagt atgatcaagc gctatgatga
     2281 gcaccaccaa gacttgactt tgctgaaggc ccttgtcaga cagcaactgc ctgagaagta
     2341 caaggaaatt ttcttcgatc agtctaaaaa tggctacgcc ggatacattg acggcggagc
     2401 aagccaggag gaattttaca aatttattaa gcccatcttg gaaaaaatgg acggcaccga
     2461 ggagctgctg gtaaagctta acagagaaga tctgttgcgc aaacagcgca ctttcgacaa
     2521 tggaagcatc ccccaccaga ttcacctggg cgaactgcac gctatcctca ggcggcaaga
     2581 ggatttctac ccctttttga aagataacag ggaaaagatt gagaaaatcc tcacatttcg
     2641 gataccctac tatgtaggcc ccctcgcccg gggaaattcc agattcgcgt ggatgactcg
     2701 caaatcagaa gagaccatca ctccctggaa cttcgaggaa gtcgtggata agggggcctc
     2761 tgcccagtcc ttcatcgaaa ggatgactaa ctttgataaa aatctgccta acgaaaaggt
     2821 gcttcctaaa cactctctgc tgtacgagta cttcacagtt tataacgagc tcaccaaggt
     2881 caaatacgtc acagaaggga tgagaaagcc agcattcctg tctggagagc agaagaaagc
     2941 tatcgtggac ctcctcttca agacgaaccg gaaagttacc gtgaaacagc tcaaagaaga
     3001 ctatttcaaa aagattgaat gtttcgactc tgttgaaatc agcggagtgg aggatcgctt
     3061 caacgcatcc ctgggaacgt atcacgatct cctgaaaatc attaaagaca aggacttcct
     3121 ggacaatgag gagaacgagg acattcttga ggacattgtc ctcaccctta cgttgtttga
     3181 agatagggag atgattgaag aacgcttgaa aacttacgct catctcttcg acgacaaagt
     3241 catgaaacag ctcaagaggc gccgatatac aggatggggg cggctgtcaa gaaaactgat
     3301 caatgggatc cgagacaagc agagtggaaa gacaatcctg gattttctta agtccgatgg
     3361 atttgccaac cggaacttca tgcagttgat ccatgatgac tctctcacct ttaaggagga
     3421 catccagaaa gcacaagttt ctggccaggg ggacagtctt cacgagcaca tcgctaatct
     3481 tgcaggtagc ccagctatca aaaagggaat actgcagacc gttaaggtcg tggatgaact
     3541 cgtcaaagta atgggaaggc ataagcccga gaatatcgtt atcgagatgg cccgagagaa
     3601 ccaaactacc cagaagggac agaagaacag tagggaaagg atgaagagga ttgaagaggg
     3661 tataaaagaa ctggggtccc aaatccttaa ggaacaccca gttgaaaaca cccagcttca
     3721 gaatgagaag ctctacctgt actacctgca gaacggcagg gacatgtacg tggatcagga
     3781 actggacatc aatcggctct ccgactacga cgtggatcat atcgtgcccc agtcttttct
     3841 caaagatgat tctattgata ataaagtgtt gacaagatcc gataaaaata gagggaagag
     3901 tgataacgtc ccctcagaag aagttgtcaa gaaaatgaaa aattattggc ggcagctgct
     3961 gaacgccaaa ctgatcacac aacggaagtt cgataatctg actaaggctg aacgaggtgg
     4021 cctgtctgag ttggataaag ccggcttcat caaaaggcag cttgttgaga cacgccagat
     4081 caccaagcac gtggcccaaa ttctcgattc acgcatgaac accaagtacg atgaaaatga
     4141 caaactgatt cgagaggtga aagttattac tctgaagtct aagctggtct cagatttcag
     4201 aaaggacttt cagttttata aggtgagaga gatcaacaat taccaccatg cgcatgatgc
     4261 ctacctgaat gcagtggtag gcactgcact tatcaaaaaa tatcccaagc ttgaatctga
     4321 atttgtttac ggagactata aagtgtacga tgttaggaaa atgatcgcaa agtctgagca
     4381 ggaaataggc aaggccaccg ctaagtactt cttttacagc aatattatga attttttcaa
     4441 gaccgagatt acactggcca atggagagat tcggaagcga ccacttatcg aaacaaacgg
     4501 agaaacagga gaaatcgtgt gggacaaggg tagggatttc gcgacagtcc ggaaggtcct
     4561 gtccatgccg caggtgaaca tcgttaaaaa gaccgaagta cagaccggag gcttctccaa
     4621 ggaaagtatc ctcccgaaaa ggaacagcga caagctgatc gcacgcaaaa aagattggga
     4681 ccccaagaaa tacggcggat tcgattctcc tacagtcgct tacagtgtac tggttgtggc
     4741 caaagtggag aaagggaagt ctaaaaaact caaaagcgtc aaggaactgc tgggcatcac
     4801 aatcatggag cgatcaagct tcgaaaaaaa ccccatcgac tttctcgagg cgaaaggata
     4861 taaagaggtc aaaaaagacc tcatcattaa gcttcccaag tactctctct ttgagcttga
     4921 aaacggccgg aaacgaatgc tcgctagtgc gggcgagctg cagaaaggta acgagctggc
     4981 actgccctct aaatacgtta atttcttgta tctggccagc cactatgaaa agctcaaagg
     5041 gtctcccgaa gataatgagc agaagcagct gttcgtggaa caacacaaac actaccttga
     5101 tgagatcatc gagcaaataa gcgaattctc caaaagagtg atcctcgccg acgctaacct
     5161 cgataaggtg ctttctgctt acaataagca cagggataag cccatcaggg agcaggcaga
     5221 aaacattatc cacttgttta ctctgaccaa cttgggcgcg cctgcagcct tcaagtactt
     5281 cgacaccacc atagacagaa agcggtacac ctctacaaag gaggtcctgg acgccacact
     5341 gattcatcag tcaattacgg ggctctatga aacaagaatc gacctctctc agctcggtgg
     5401 agacagcagg gctgacccca agaagaagag gaaggtgggt ggaggaggta ccggcggtgg
     5461 aggctcagca gaatacgtac gagctctgtt tgacttcaat gggaatgacg aggaggatct
     5521 cccctttaag aagggcgata ttctccgcat cagagataag cccgaagaac aatggtggaa
     5581 tgccgaggat agcgaaggga aaaggggcat gattctggtg ccatatgtgg agaaatattc
     5641 cggtgactac aaagaccatg atggggatta caaagaccac gacatcgact acaaagacga
     5701 cgacgataaa tcagggggtc ccaagaagaa aagaaaagtc ggtaccatga ccaacctttc
     5761 cgacatcata gagaaggaaa caggcaaaca gttggtcatc caagagtcga tactcatgct
     5821 tcctgaagaa gttgaggagg tcattgggaa taagccggaa agtgacattc tcgtacacac
     5881 tgcgtatgat gagagcaccg atgagaacgt gatgctgctc acgtcagatg ccccagagta
     5941 caaaccctgg gctctggtga ttcaggactc taatggagag aacaagatca agatgctagg
     6001 aggcggtgga agcggcgcaa caaacttctc tctgctgaaa caagccggag atgtcgaaga
     6061 gaatcctgga ccgatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     6121 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     6181 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     6241 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     6301 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     6361 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     6421 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     6481 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     6541 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     6601 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     6661 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     6721 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     6781 gctgtacaag tgaagcggcc gcactcctca ggtgcaggct gcctatcaga aggtggtggc
     6841 tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa
     6901 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     6961 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     7021 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     7081 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
     7141 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     7201 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     7261 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     7321 gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     7381 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     7441 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7501 tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt
     7561 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     7621 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     7681 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg
     7741 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     7801 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     7861 gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7921 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7981 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     8041 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     8101 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     8161 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     8221 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     8281 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     8341 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     8401 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     8461 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     8521 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     8581 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     8641 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     8701 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     8761 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     8821 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8881 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8941 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     9001 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     9061 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     9121 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     9181 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     9241 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     9301 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     9361 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     9421 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     9481 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     9541 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     9601 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     9661 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     9721 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     9781 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9841 cccgaaaagt gccacctggg tcgacattga ttattgacta gttattaata gtaatcaatt
     9901 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     9961 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
    10021 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
    10081 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
    10141 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
    10201 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
    10261 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
    10321 ttttaattat tttgtgcagc gatgggggcg ggggggggg
//
