LOCUS       sequence-232775-       12714 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 12714)
  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 12714)
  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..12714
                     /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          945..1962
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1886..1908
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1970..1989
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(2066..2099)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2228..2362
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2465..2599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2702..2836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2860..2893)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2912..2921
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2921..2986
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2993..3013
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3038..3061,3062..3064,3065..7138)
                     /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             3038..7138
                     /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             7139..7186
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    7208..7432
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    7460..7493
                     /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        7557..8056
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7592..7615)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7989..8008
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        8068..8115
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8134..8937
                     /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(8197..8216)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8807..8826
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8975..9199
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    9254..9287
                     /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(9939..9958)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9940..9958)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9968..9985)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9968..9984)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9977..9999)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10126..10581
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10213..10232)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10423..10444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10607..10711
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10712..10780,10781..11572)
                     /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(10930..10949)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11743..12331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12232..12251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12485..12502
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12619..12640
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12655..12660,12661..12678,12679..12685)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12693..12709
                     /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(12698..12714,1..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 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag
      841 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
      901 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcggggagtc gctgcgacgc
      961 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
     1021 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     1081 cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc
     1141 cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg
     1201 gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
     1261 gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg
     1321 tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga
     1381 gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt
     1441 gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc
     1501 gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc
     1561 ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg
     1621 gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg
     1681 tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc
     1741 ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
     1801 ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc
     1861 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga
     1921 gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
     1981 gttattgtgc tgtctcatca ttttggcaaa gaattgattt gataccgcgg gccctcgaca
     2041 ctagtgaacc tcttcgaggg acctaataac ttcgtatagc atacattata cgaagttata
     2101 ttaagggttc cggatccaca tgatcataat cagccatacc acatttgtag aggttttact
     2161 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2221 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2281 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2341 tgtatcttat catgtctgga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2401 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2461 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2521 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2581 atcttatcat gtctggatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2641 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2701 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2761 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2821 ttatcatgtc tggatctgtc gacctgcagc caagctagta taacttcgta tagcatacat
     2881 tatacgaagt tattgcggcc gcggactcgc cgccaccatg gactataagg accacgacgg
     2941 agactacaag gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa
     3001 gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct
     3061 ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
     3121 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
     3181 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
     3241 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     3301 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     3361 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     3421 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     3481 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     3541 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     3601 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     3661 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     3721 gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac
     3781 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     3841 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     3901 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     3961 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     4021 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     4081 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     4141 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     4201 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     4261 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     4321 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     4381 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     4441 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     4501 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     4561 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     4621 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt
     4681 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     4741 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     4801 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     4861 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     4921 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     4981 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     5041 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     5101 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     5161 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     5221 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     5281 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     5341 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     5401 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     5461 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     5521 catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga
     5581 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa
     5641 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc
     5701 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag
     5761 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     5821 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct
     5881 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     5941 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     6001 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
     6061 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     6121 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     6181 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
     6241 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat
     6301 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
     6361 tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa
     6421 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
     6481 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
     6541 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
     6601 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
     6661 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
     6721 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
     6781 cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat
     6841 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
     6901 agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat
     6961 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
     7021 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca
     7081 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa
     7141 aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaagtgat aaggcctcga
     7201 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     7261 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     7321 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     7381 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     7441 cataggatcc ccgaattccg aagttcctat tctctagaaa gtataggaac ttcaggtctg
     7501 aagaggagtt tacgtccagc caagctagct tggctgcagg tcgtcgaaat tctaccgggt
     7561 aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac
     7621 ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca
     7681 accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa
     7741 gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct
     7801 cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg
     7861 cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg
     7921 gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg
     7981 aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc
     8041 atctccgggc ctttcgacct gcagcctgtt gacaattaat catcggcata gtatatcggc
     8101 atagtataat acgacaaggt gaggaactaa accatgggat cggccattga acaagatgga
     8161 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     8221 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     8281 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     8341 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     8401 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     8461 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     8521 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     8581 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     8641 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgatga tctcgtcgtg
     8701 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     8761 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     8821 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     8881 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaggg
     8941 gatcaattct ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     9001 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     9061 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     9121 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     9181 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc gactagagct
     9241 tgcggaaccc ttcgaagttc ctattctcta gaaagtatag gaacttcatc agtcaggtac
     9301 ataatataac ttcgcggacc gggagtcttc tgggcaggct taaaggctaa cctggtgcgt
     9361 ggggcgttgt cctgcagagg aattgaacag gtgtaaaatt ggaggggcaa gacttcccac
     9421 agattttcga ttgtgttgtt aagtattgta ataggggcaa ataagggaaa tagactaggc
     9481 actcacctgg ggttttatgc agcaaaacta caggttatta ttgcttgtga tccgccctgg
     9541 agaatttttc accgaggtag attgaagaca tgcccaccca aattttaata ttcttccact
     9601 tgcgatcctt gctacagtat gaaattacag tatcgtgaat tagaatatat aagcagaatt
     9661 ttaagcattt taaaagagcc cagtacttca tgtctgtctc tcccacttct gcagccctat
     9721 caaagggtat tttagcacac tcattttagt cccattttca tttgttgtac tggcttatcc
     9781 aatccctaga cagagcactg gcattccctc tctcctgatc ttagaagtcc gatgactcat
     9841 gaaaccagac agattagttt catacaccac aaatcgaggc tgtagctggg gcctttaaca
     9901 ttgcagtttt tttattcttc agtacggtac ccaattcgcc ctatagtgag tcgtattacg
     9961 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
    10021 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
    10081 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
    10141 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
    10201 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
    10261 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
    10321 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
    10381 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
    10441 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
    10501 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
    10561 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
    10621 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
    10681 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
    10741 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
    10801 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
    10861 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
    10921 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
    10981 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
    11041 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
    11101 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
    11161 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
    11221 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
    11281 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
    11341 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
    11401 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
    11461 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
    11521 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
    11581 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
    11641 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
    11701 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
    11761 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
    11821 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
    11881 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
    11941 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
    12001 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
    12061 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
    12121 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
    12181 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
    12241 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
    12301 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
    12361 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
    12421 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
    12481 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
    12541 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
    12601 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
    12661 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttca
//
