LOCUS       sequence-363084-        9253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of spCas9, gRNA targeting the end of Tubb3 gene and donor
            GFP(Y39TAG). Can be used for amber codon suppression and click 
            chemistry labeling of endogenous beta-3 tubulin in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9253)
  AUTHORS   Arsic A, Hagemann C, Stajkovic N, Schubert T, Nikic-Spiegel I
  TITLE     Minimal genetically encoded tags for fluorescent protein labeling in
            living neurons.
  JOURNAL   Nat Commun. 2022 Jan 14;13(1):314. doi: 10.1038/s41467-022-27956-y.
  PUBMED    35031604
REFERENCE   2  (bases 1 to 9253)
  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 
            Commun."; date: "2022-01-14"; volume: "13(1):314. doi"; pages: " 
            10.1038/s41467-022-27956-y"
COMMENT     Sequence Label: pORANGE_Tubb3 GFP(Y39TAG) KI
FEATURES             Location/Qualifiers
     source          1..9253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(42..44,45..758)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDAT*GKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(345..364)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     692..713
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        828..1113
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          1393..1620
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1641..1667
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1671..1691
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1701..5801
                     /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             1701..5801
                     /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             5805..5825
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5865..5882)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5871..6078
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6087..6227
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6087..6216
                     /label=AAV2 ITR
     rep_origin      6302..6757
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6389..6408)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6599..6620
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6774..6793)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6893..6915
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6953..6971)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7039..7143
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7144..7212,7213..8004)
                     /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(7362..7381)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8175..8763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8664..8683
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8825..9073
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8825..8845
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8996..9015
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9093..9168
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 ctagaagctt gctgcgagca acttcacttg ggggatcagg cgtgagcaag ggcgaggagc
       61 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      121 tcagcgtgtc cggcgagggc gagggcgatg ccacctaggg caagctgacc ctgaagttca
      181 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
      241 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      301 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      361 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      421 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
      481 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcc aacttcaaga
      541 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
      601 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
      661 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
      721 ccgggatcac tctcggcatg gacgagctgt acaagtaatg tgctgcgagc aacttcactt
      781 gggctcgagg atcctaggac gcgtcctagg acgcgtccag aggtacccgt tacataactt
      841 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaatagta
      901 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      961 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     1021 aaatggcccg cctggcattg tgcccagtac atgaccttat gggactttcc tacttggcag
     1081 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
     1141 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
     1201 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccrggsgg ggsggggsgg
     1261 ggsgrggggs ggggsggggs grggcggrga ggkgcggcgg cagccaatca gagcggcgcg
     1321 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
     1381 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
     1441 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
     1501 ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg
     1561 gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag
     1621 gttggaccgg tgccaccatg tacccatacg atgttccaga ttacgcttcg ccgaagaaaa
     1681 agcgcaaggt cgaagcgtcc gacaagaagt acagcatcgg cctggacatc ggcaccaact
     1741 ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc
     1801 tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca
     1861 gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac
     1921 ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg
     1981 acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc
     2041 ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca
     2101 tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct
     2161 atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga
     2221 accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc
     2281 tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca
     2341 gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga
     2401 atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca
     2461 acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc
     2521 tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga
     2581 acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg
     2641 cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc
     2701 tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga
     2761 gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt
     2821 tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca
     2881 gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc
     2941 acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg
     3001 acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc
     3061 tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc
     3121 cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga
     3181 tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt
     3241 acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga
     3301 gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga
     3361 ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct
     3421 tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc
     3481 acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca
     3541 ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac
     3601 ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga
     3661 gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt
     3721 ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc
     3781 agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg
     3841 gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga
     3901 agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca
     3961 agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga
     4021 agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga
     4081 tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact
     4141 acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg
     4201 actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca
     4261 aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg
     4321 tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga
     4381 gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg
     4441 gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc
     4501 tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag
     4561 tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag
     4621 tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa
     4681 ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg
     4741 tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca
     4801 agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg
     4861 gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg
     4921 ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg
     4981 tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga
     5041 acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg
     5101 acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca
     5161 agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg
     5221 agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga
     5281 tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg
     5341 cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact
     5401 tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga
     5461 aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg
     5521 agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca
     5581 acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc
     5641 tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga
     5701 ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc
     5761 tgtacgagac acggatcgac ctgtctcagc tgggaggcga cagccccaag aagaagagaa
     5821 aggtggaggc cagctaagaa ttctaactag agctcgctga tcagcctcga ctgtgccttc
     5881 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     5941 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     6001 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagagaa
     6061 tagcaggcat gctggggagc ggccgcagga acccctagtg atggagttgg ccactccctc
     6121 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     6181 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     6241 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     6301 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6361 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6421 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6481 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     6541 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6601 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     6661 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     6721 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     6781 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     6841 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     6901 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     6961 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     7021 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     7081 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     7141 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     7201 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     7261 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7321 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     7381 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7441 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7501 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7561 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7621 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7681 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7741 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7801 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga
     7861 agccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7921 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7981 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     8041 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     8101 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8161 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8221 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8281 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     8341 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8401 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8461 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8521 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8581 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8641 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8701 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8761 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     8821 atgtgagggc ctatttccca tgattccttc atatttgcat atacgataca aggctgttag
     8881 agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg
     8941 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
     9001 tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa
     9061 aggacgaaac accgctgcga gcaacttcac ttgttttaga gctagaaata gcaagttaaa
     9121 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttgtttta
     9181 gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc gccaattctg
     9241 cagacaaatg gct
//
