LOCUS       sequence-239344-        8535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV promoter expression plasmid for codon-optimized bpNLS-32AA 
            linker-hSpCas9n(D10A)-bpNLS-P2A-EGFP (ABEmax control without TadA 
            domains)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8535)
  AUTHORS   Grunewald J, Zhou R, Garcia SP, Iyer S, Lareau CA, Aryee MJ, Joung 
            JK
  TITLE     Transcriptome-wide off-target RNA editing induced by CRISPR-guided 
            DNA base editors.
  JOURNAL   Nature. 2019 Apr 17. pii: 10.1038/s41586-019-1161-z. doi: 
            10.1038/s41586-019-1161-z.
  PUBMED    30995674
REFERENCE   2  (bases 1 to 8535)
  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: "Nature. 
            2019 Apr 17. pii: 10.1038/s41586-019-1161-z. doi: 
            10.1038/s41586-019-1161-z."
COMMENT     Sequence Label: NLS-nCas9-NLS-P2A-EGFP (pRZ70)
FEATURES             Location/Qualifiers
     source          1..8535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             445..465
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(562..585,586..588,589..4662)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             562..4662
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4705..4725
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4735..4791
                     /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(4792..4794,4795..4797,4798..5508)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4792..4794,4795..4797,4798..5508)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5098..5117)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5445..5466
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5521..5541
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5550..5567
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5590..5607)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5596..5820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5891..5907)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5891..5907)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5904..5926)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5915..5931
                     /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(5939..5945,5946..5963,5964..5969))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5984..6005
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6122..6139)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6293..6881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6373..6392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7052..7843,7844..7912))
                     /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     7675..7694
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7913..8017)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8096..8115)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(8287..8535,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca
      421 gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctccgg aggatctagc
      481 ggaggctcct ctggctctga gacacctggc acaagcgaga gcgcaacacc tgaaagcagc
      541 gggggcagca gcggggggtc agacaagaag tacagcatcg gcctggccat cggcaccaac
      601 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
      661 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
      721 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
      781 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
      841 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
      901 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
      961 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     1021 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     1081 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     1141 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     1201 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     1261 aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     1321 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     1381 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     1441 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     1501 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     1561 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     1621 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     1681 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     1741 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     1801 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     1861 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     1921 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     1981 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     2041 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     2101 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     2161 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     2221 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     2281 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     2341 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     2401 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     2461 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     2521 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     2581 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     2641 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     2701 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     2761 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     2821 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     2881 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     2941 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     3001 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     3061 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
     3121 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     3181 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     3241 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     3301 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     3361 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     3421 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     3481 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     3541 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     3601 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     3661 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     3721 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     3781 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     3841 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     3901 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     3961 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     4021 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     4081 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     4141 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     4201 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     4261 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     4321 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
     4381 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
     4441 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
     4501 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
     4561 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
     4621 ctgtacgaga cacggatcga cctgtctcag ctgggaggtg actctggcgg ctcaaaaaga
     4681 accgccgacg gcagcgaatt cgagcccaag aagaagagga aagtcggaag cggagctact
     4741 aacttcagcc tgctgaagca ggctggagac gtggaggaga accctggacc tatggtgagc
     4801 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     4861 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     4921 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     4981 accctgacct atggagtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     5041 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     5101 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     5161 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     5221 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     5281 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     5341 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     5401 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     5461 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc tggtggttct
     5521 cccaagaaga agaggaaagt ctaaccggtc atcatcacca tcaccattga gtttaaaccc
     5581 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     5641 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     5701 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     5761 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     5821 cttctgaggc ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg
     5881 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5941 acatacgagc cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca
     6001 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6061 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6121 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6181 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6241 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6301 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6361 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6421 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6481 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6541 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6601 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6661 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6721 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6781 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6841 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6901 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6961 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7021 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7081 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7141 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7201 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7261 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7321 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7381 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7441 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7501 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7561 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7621 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7681 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7741 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7801 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7861 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7921 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7981 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8041 ctgacgtcga cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa
     8101 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     8161 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     8221 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     8281 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     8341 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     8401 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     8461 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     8521 tacatcaagt gtatc
//
