LOCUS       sequence-455783-        9644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing Cas9 and gRNAs for mouse Ift88 and Pkd2. Use for 
            disruption of mouse Ift88 and Pkd2 in cultured cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9644)
  TITLE     Plasmid expressing Cas9 and 2 gRNAs for mouse Pkd2 and Ift88
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9644)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCJ1023
FEATURES             Location/Qualifiers
     source          1..9644
                     /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        270..345
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        434..682
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        703..778
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        796..1081
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1083..1360
                     /label=chicken beta-actin promoter
     intron          1361..1588
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1600..1609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1609..1674
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1681..1701
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1726..5826
                     /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             1726..5826
                     /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             5827..5874
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5890..5943
                     /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(5944..5946,5947..6657)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5944..5946,5947..6657)
                     /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(6247..6266)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6594..6615
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6685..6702)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6691..6898
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6907..7047
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6907..7036
                     /label=AAV2 ITR
     rep_origin      7122..7577
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7209..7228)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7419..7440
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7594..7613)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7713..7735
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7773..7791)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7859..7963
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7964..8032,8033..8824)
                     /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(8182..8201)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8995..9583
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9484..9503
                     /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 cgaaacacct caatgggaag accgatgacg ttttagagct agaaatagca agttaaaata
      301 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag
      361 ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag
      421 acaaatggct ctagagggcc tatttcccat gattccttca tatttgcata tacgatacaa
      481 ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa
      541 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
      601 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
      661 cttgtggaaa ggacgaaaca ccacgcacgc agttccattc cggttttaga gctagaaata
      721 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
      781 tttttctagg gtacccgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      841 cgacccccgc ccattgacgt caatagtaac gccaataggg actttccatt gacgtcaatg
      901 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      961 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattgtg cccagtacat
     1021 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1081 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc
     1141 aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg
     1201 ggggcgcgcg ccgggggggg gggggggggg gggggggggg ggsggggsga ggsggagagg
     1261 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
     1321 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
     1381 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
     1441 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagctgag
     1501 caagaggtaa gggtttaagg gatggttggt tggtggggta ttaatgttta attacctgga
     1561 gcacctgcct gaaatcactt tttttcaggt tggaccggtg ccaccatgga ctataaggac
     1621 cacgacggag actacaagga tcatgatatt gattacaaag acgatgacga taagatggcc
     1681 ccaaagaaga agcggaaggt cggtatccac ggagtcccag cagccgacaa gaagtacagc
     1741 atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
     1801 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
     1861 ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga
     1921 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
     1981 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
     2041 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     2101 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     2161 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     2221 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     2281 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     2341 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     2401 gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg
     2461 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     2521 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     2581 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     2641 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     2701 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     2761 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga
     2821 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     2881 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     2941 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     3001 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     3061 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     3121 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct
     3181 tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     3241 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa
     3301 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag
     3361 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     3421 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     3481 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     3541 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     3601 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     3661 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     3721 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     3781 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     3841 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     3901 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     3961 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     4021 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     4081 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     4141 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     4201 gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt
     4261 ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag
     4321 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg
     4381 ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc
     4441 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     4501 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat
     4561 gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     4621 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     4681 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     4741 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     4801 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     4861 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     4921 ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg
     4981 ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     5041 aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg
     5101 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     5161 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     5221 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     5281 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     5341 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     5401 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     5461 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg
     5521 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     5581 ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc
     5641 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     5701 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     5761 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
     5821 ggcgacaaaa ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaaggaattc
     5881 ggcagtggag agggcagagg aagtctgcta acatgcggtg acgtcgagga gaatcctggc
     5941 ccagtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     6001 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     6061 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     6121 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     6181 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     6241 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     6301 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     6361 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     6421 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     6481 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     6541 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     6601 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggaa
     6661 ttctaactag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     6721 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     6781 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     6841 gggtggggca ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc
     6901 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     6961 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     7021 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     7081 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     7141 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     7201 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     7261 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     7321 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     7381 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     7441 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     7501 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     7561 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7621 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     7681 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     7741 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     7801 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     7861 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7921 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7981 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     8041 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     8101 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     8161 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     8221 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     8281 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     8341 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     8401 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     8461 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     8521 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     8581 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     8641 ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc
     8701 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8761 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8821 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     8881 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8941 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     9001 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     9061 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     9121 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     9181 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     9241 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     9301 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     9361 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     9421 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     9481 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     9541 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     9601 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
