LOCUS       sequence-238032-       12710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A donor plasmid with homologous arms matching rat Rosa26 and 
            expressing a Cre-dependent FLAG-tagged SpCas9n and a NeoR selectable
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12710)
  AUTHORS   Back S, Necarsulmer J, Whitaker LR, Coke LM, Koivula P, Heathward 
            EJ, Fortuno LV, Zhang Y, Yeh CG, Baldwin HA, Spencer MD, 
            Mejias-Aponte CA, Pickel J, Hoffman AF, Spivak CE, Lupica CR, 
            Underhill SM, Amara SG, Domanskyi A, Anttila JE, Airavaara M, Hope 
            BT, Hamra FK, Richie CT, Harvey BK
  TITLE     Neuron-Specific Genome Modification in the Adult Rat Brain Using 
            CRISPR-Cas9 Transgenic Rats.
  JOURNAL   Neuron. 2019 Feb 8. pii: S0896-6273(19)30062-5. doi: 
            10.1016/j.neuron.2019.01.035.
  PUBMED    30792150
REFERENCE   2  (bases 1 to 12710)
  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: "Neuron. 
            2019 Feb 8. pii: S0896-6273(19)30062-5. doi: 
            10.1016/j.neuron.2019.01.035."
COMMENT     Sequence Label: pOTTC1076 - prRosa26v1 LSL FLAG-SpCas9n NeoR
FEATURES             Location/Qualifiers
     source          1..12710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..19
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3..19
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     38..58
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        40..58
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        361..664
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        666..943
                     /label=chicken beta-actin promoter
     intron          944..1960
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1884..1906
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1968..1987
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(2062..2095)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2224..2358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2461..2595
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2698..2832
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2856..2889)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2908..2917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2917..2982
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2989..3009
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3034..3057,3058..3060,3061..7134)
                     /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             3034..7134
                     /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             7135..7182
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    7204..7428
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    7456..7489
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        7553..8052
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7588..7611)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7985..8004
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        8064..8111
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8130..8933
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8193..8212)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8803..8822
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8971..9195
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    9250..9283
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(9935..9954)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9936..9954)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9964..9981)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9964..9980)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9973..9995)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10122..10577
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10209..10228)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10419..10440
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10603..10707
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10708..10776,10777..11568)
                     /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(10926..10945)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11739..12327
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12228..12247
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12481..12498
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12615..12636
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12651..12656,12657..12674,12675..12681)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12689..12705
                     /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)."
     primer_bind     join(12694..12710,1..6)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
       61 acaaaagctg gagctcttgg agacaagaag cacttgctct ccaaaagtcg gtttgagtta
      121 tcattaaggg agctgcagtg gagtaggcgg agaaaaggcc gcacccttct caggacgggg
      181 gaggggagtg ttgcaatacc tttctgggag ttctctgctg cctcctgtct tctgaggacc
      241 gccctgggcc tggaagattc ccttcccacg cgtaaccgtt taaacaattc tgcaggaatc
      301 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      361 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      421 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      481 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      541 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      601 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      661 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
      721 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg
      781 gggggggcgc gcgccrggsg gggsggggsg gggsgrgggg sggggsgggg sgrggcggag
      841 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
      901 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg
      961 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
     1021 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
     1081 cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg
     1141 ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg
     1201 gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg
     1261 gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg
     1321 cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
     1381 agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc
     1441 tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt
     1501 gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg
     1561 ggrgggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc
     1621 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
     1681 ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg
     1741 ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc
     1801 gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct
     1861 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc
     1921 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     1981 tattgtgctg tctcatcatt ttggcaaaga attgatttga taccgcgggc tcgacactag
     2041 tgaacctctt cgagggacct aataacttcg tatagcatac attatacgaa gttatattaa
     2101 gggttccgga tccacatgat cataatcagc cataccacat ttgtagaggt tttacttgct
     2161 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2221 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2281 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2341 tcttatcatg tctggatcat aatcagccat accacatttg tagaggtttt acttgcttta
     2401 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2461 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2521 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2581 tatcatgtct ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2641 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2701 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2761 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     2821 catgtctgga tctgtcgacc tgcagccaag ctagtataac ttcgtatagc atacattata
     2881 cgaagttatt gcggccgcgg actcgccgcc accatggact ataaggacca cgacggagac
     2941 tacaaggatc atgatattga ttacaaagac gatgacgata agatggcccc aaagaagaag
     3001 cggaaggtcg gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggcc
     3061 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
     3121 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
     3181 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
     3241 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
     3301 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
     3361 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
     3421 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
     3481 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
     3541 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
     3601 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     3661 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     3721 ggcgagaaga agaatggcct gttcggcaac ctgattgccc tgagcctggg cctgaccccc
     3781 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     3841 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     3901 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     3961 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     4021 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     4081 ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa
     4141 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     4201 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     4261 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     4321 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     4381 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     4441 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc
     4501 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     4561 cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg
     4621 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac
     4681 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
     4741 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
     4801 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
     4861 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
     4921 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
     4981 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
     5041 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
     5101 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
     5161 gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc
     5221 cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg
     5281 atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc
     5341 cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag
     5401 ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag
     5461 ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc
     5521 aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac
     5581 tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg
     5641 ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag
     5701 ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa
     5761 ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac
     5821 gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc
     5881 cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc
     5941 cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac
     6001 gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac
     6061 ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc
     6121 aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt
     6181 accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg
     6241 gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc
     6301 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
     6361 ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag
     6421 tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
     6481 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
     6541 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
     6601 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
     6661 aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc
     6721 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
     6781 gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc
     6841 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg
     6901 ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc
     6961 cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc
     7021 atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag
     7081 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaaaagg
     7141 ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa agtgataagg cctcgagcct
     7201 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     7261 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     7321 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     7381 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgagcata
     7441 ggatccccga attccgaagt tcctattctc tagaaagtat aggaacttca ggtctgaaga
     7501 ggagtttacg tccagccaag ctagcttggc tgcaggtcgt cgaaattcta ccgggtaggg
     7561 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg
     7621 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg
     7681 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc
     7741 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact
     7801 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc
     7861 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc
     7921 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc
     7981 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct
     8041 ccgggccttt cgacctgcag cctgttgaca attaatcatc ggcatagtat atcggcatag
     8101 tataatacga caaggtgagg aactaaacca tgggatcggc cattgaacaa gatggattgc
     8161 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     8221 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     8281 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     8341 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     8401 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     8461 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     8521 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     8581 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     8641 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgatgatctc gtcgtgaccc
     8701 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     8761 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     8821 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     8881 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaggggatc
     8941 aattctctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     9001 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     9061 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     9121 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     9181 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgact agagcttgcg
     9241 gaacccttcg aagttcctat tctctagaaa gtataggaac ttcatcagtc aggtacataa
     9301 tataacttcg cggaccggga gtcttctggg caggcttaaa ggctaacctg gtgcgtgggg
     9361 cgttgtcctg cagaggaatt gaacaggtgt aaaattggag gggcaagact tcccacagat
     9421 tttcgattgt gttgttaagt attgtaatag gggcaaataa gggaaataga ctaggcactc
     9481 acctggggtt ttatgcagca aaactacagg ttattattgc ttgtgatccg ccctggagaa
     9541 tttttcaccg aggtagattg aagacatgcc cacccaaatt ttaatattct tccacttgcg
     9601 atccttgcta cagtatgaaa ttacagtatc gtgaattaga atatataagc agaattttaa
     9661 gcattttaaa agagcccagt acttcatgtc tgtctctccc acttctgcag ccctatcaaa
     9721 gggtatttta gcacactcat tttagtccca ttttcatttg ttgtactggc ttatccaatc
     9781 cctagacaga gcactggcat tccctctctc ctgatcttag aagtccgatg actcatgaaa
     9841 ccagacagat tagtttcata caccacaaat cgaggctgta gctggggcct ttaacattgc
     9901 agttttttta ttcttcagta cggtacccaa ttcgccctat agtgagtcgt attacgcgcg
     9961 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
    10021 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
    10081 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
    10141 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
    10201 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
    10261 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
    10321 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
    10381 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
    10441 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
    10501 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
    10561 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
    10621 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
    10681 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
    10741 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
    10801 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
    10861 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
    10921 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
    10981 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
    11041 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
    11101 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
    11161 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
    11221 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
    11281 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
    11341 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
    11401 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
    11461 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
    11521 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
    11581 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
    11641 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
    11701 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
    11761 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
    11821 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
    11881 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
    11941 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
    12001 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
    12061 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
    12121 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
    12181 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
    12241 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
    12301 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
    12361 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
    12421 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
    12481 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
    12541 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
    12601 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
    12661 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
//
