LOCUS       sequence-232772-       12060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A donor plasmid with homologous arms matching rat Rosa26 and 
            expressing a Cre-dependent FLAG-tagged SpCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12060)
  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 12060)
  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: pOTTC1080 - prRosa26v2 LSL FLAG-SpCas9
FEATURES             Location/Qualifiers
     source          1..12060
                     /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        1191..1494
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1496..1773
                     /label=chicken beta-actin promoter
     intron          1775..2792
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2716..2738
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2800..2819
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(2896..2929)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3058..3192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3295..3429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3532..3666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3690..3723)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3742..3751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3751..3816
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3823..3843
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3868..7968
                     /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             3868..7968
                     /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             7969..8016
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(8032..8049)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8038..8262
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9285..9304)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9286..9304)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9314..9331)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9314..9330)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9323..9345)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9472..9927
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9559..9578)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9769..9790
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9953..10057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10058..10126,10127..10918)
                     /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(10276..10295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11089..11677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11578..11597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11831..11848
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11965..11986
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12001..12006,12007..12024,12025..12031)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12039..12055
                     /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(12044..12060,1..6)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
       61 acaaaagctg gagctcgccc tccaaggacg gtggagccgc tttgtgggac agctgggttc
      121 gattcgttaa ccctggaagg ggcaagcggg tggtagtcag gaatccggcc gccctgcagc
      181 aaccggaggg ggagggagaa gggagcggaa aagtctccac cggacgcggc catggctccc
      241 acggggggcg gagaagcgct tccggtcgat gtctcatcgc tgattggctg cttttcctcc
      301 cgccgcgtgt gaaaacacaa atggcgtgtt ttggttggag tgaggcgcct gtcaattaac
      361 ggctgccgga gtgcgcagcc gctgactgcc tcgctgtgcc cactgggtgg ggcgggaggt
      421 aggtggggtg aggcgagctg gacgtgcggg cgcggtcggc ctctggcggg gcgggggagg
      481 ggagggtcag cgaaagtggc tggcgcgtga gcggcctccc accctcccct tcctctgggg
      541 gagtcgtttt acccgccgcc ggcctggcct cgtcatctga ttggctctcg gggctcagaa
      601 aactggcctt tgcaattggc ccgcgttcat gcaagttcag tccctaagct ggctggcggg
      661 ggcggcaggg aggcgctcac aggttccggc cctcccccca ggccccgcgc cgcagagtct
      721 ggccccgcgc ccctgcgcaa cgtggcagga agcgcgcgct gggggcgggg acgggcggtc
      781 ggtctgagcg gcgggcgggt gcaaacggga ttcctccttg agttgtggca ctgaggaacg
      841 tgctgaacaa gacctacatt gcactccagg gagtggatga aggagttggg gctcagtcgg
      901 gttgtattgg agacaagaag cacttgctct ccaaaagtcg gtttgagtta tcattaaggg
      961 agctgcagtg gagtaggcgg agaaaaggcc gcacccttct caggacgggg gaggggagtg
     1021 ttgcaatacc tttctgggag ttctctgctg cctcctgtct tctgaggacc gccctgggcc
     1081 tggaagattc ccttcccacg cgtaaccgtt taaacaattc tgcaggaatc tagttattaa
     1141 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     1201 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     1261 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     1321 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     1381 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     1441 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
     1501 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
     1561 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc
     1621 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
     1681 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
     1741 cggccctata aaaagcgaag cgcgcggcgg gcggggagtc gctgcgacgc tgccttcgcc
     1801 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
     1861 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
     1921 aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc
     1981 ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg
     2041 cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg
     2101 cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
     2161 gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg
     2221 tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg
     2281 gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg
     2341 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
     2401 cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
     2461 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
     2521 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     2581 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     2641 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
     2701 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     2761 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     2821 tgtctcatca ttttggcaaa gaattgattt gataccgcgg gccctcgaca ctagtgaacc
     2881 tcttcgaggg acctaataac ttcgtatagc atacattata cgaagttata ttaagggttc
     2941 cggatccaca tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3001 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3061 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3121 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3181 catgtctgga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3241 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3301 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3361 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3421 gtctggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3481 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3541 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3601 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3661 tggatctgtc gacctgcagc caagctagta taacttcgta tagcatacat tatacgaagt
     3721 tattgcggcc gcggactcgc cgccaccatg gactataagg accacgacgg agactacaag
     3781 gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag
     3841 gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc
     3901 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     3961 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     4021 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     4081 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     4141 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     4201 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     4261 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     4321 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     4381 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     4441 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     4501 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     4561 aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc
     4621 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     4681 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     4741 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     4801 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     4861 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     4921 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     4981 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     5041 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     5101 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     5161 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     5221 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     5281 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     5341 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     5401 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     5461 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     5521 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     5581 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     5641 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     5701 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     5761 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     5821 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     5881 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     5941 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     6001 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     6061 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     6121 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     6181 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     6241 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     6301 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     6361 ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc
     6421 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     6481 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     6541 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     6601 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     6661 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     6721 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     6781 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     6841 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
     6901 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     6961 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     7021 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
     7081 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     7141 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     7201 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     7261 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     7321 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     7381 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     7441 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     7501 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     7561 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     7621 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     7681 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     7741 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     7801 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
     7861 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     7921 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg
     7981 gccacgaaaa aggccggcca ggcaaaaaag aaaaagtgat aaggcctcga gcctcgactg
     8041 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     8101 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     8161 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     8221 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag cataggatcc
     8281 ccgaattcga gtcttctggg caggcttaaa ggctaacctg gtgcgtgggg cgttgtcctg
     8341 cagaggaatt gaacaggtgt aaaattggag gggcaagact tcccacagat tttcgattgt
     8401 gttgttaagt attgtaatag gggcaaataa gggaaataga ctaggcactc acctggggtt
     8461 ttatgcagca aaactacagg ttattattgc ttgtgatccg ccctggagaa tttttcaccg
     8521 aggtagattg aagacatgcc cacccaaatt ttaatattct tccacttgcg atccttgcta
     8581 cagtatgaaa ttacagtatc gtgaattaga atatataagc agaattttaa gcattttaaa
     8641 agagcccagt acttcatgtc tgtctctccc acttctgcag ccctatcaaa gggtatttta
     8701 gcacactcat tttagtccca ttttcatttg ttgtactggc ttatccaatc cctagacaga
     8761 gcactggcat tccctctctc ctgatcttag aagtccgatg actcatgaaa ccagacagat
     8821 tagtttcata caccacaaat cgaggctgta gctggggcct ttaacattgc agttttttta
     8881 ttcttcagta cactttgttg attctttgcc ttgatcttga cttcaggttc tatcaccacc
     8941 ccctcagatg gtgttccaca cttgggccta ttcacagttc agagagcttt acaacaatag
     9001 atgtattgag aatccaacct aaagttcagc tttttactcc catgaatgcc tctttccttt
     9061 ttctccattt ataaactgag ccatttcctg ttaatggttt acagatgaat atctcctccc
     9121 ccaatatcac ctgatgtatc ttacattttg ccaggcttag attgtcttaa aaggtacata
     9181 aattaacatg tgaaatttac tccttaatgc ttcagtggat ttcatgagtg cagtacagaa
     9241 gactggtaat gggctaataa cttttatttc aggtacccaa ttcgccctat agtgagtcgt
     9301 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     9361 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     9421 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     9481 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     9541 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     9601 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     9661 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     9721 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     9781 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     9841 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     9901 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     9961 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
    10021 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
    10081 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
    10141 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
    10201 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
    10261 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
    10321 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
    10381 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
    10441 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
    10501 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
    10561 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
    10621 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
    10681 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
    10741 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
    10801 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
    10861 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
    10921 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
    10981 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
    11041 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
    11101 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
    11161 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
    11221 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
    11281 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
    11341 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
    11401 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
    11461 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
    11521 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
    11581 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
    11641 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
    11701 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
    11761 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
    11821 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
    11881 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
    11941 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
    12001 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
//
