LOCUS       sequence-239523-       12056 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 12056)
  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 12056)
  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..12056
                     /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          1774..2790
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2714..2736
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2798..2817
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(2892..2925)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3054..3188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3291..3425
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3528..3662
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3686..3719)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3738..3747
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3747..3812
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3819..3839
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3864..7964
                     /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             3864..7964
                     /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             7965..8012
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(8028..8045)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8034..8258
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9281..9300)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9282..9300)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9310..9327)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9310..9326)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9319..9341)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9468..9923
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9555..9574)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9765..9786
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9949..10053
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10054..10122,10123..10914)
                     /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(10272..10291)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11085..11673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11574..11593
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11827..11844
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11961..11982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11997..12002,12003..12020,12021..12027)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12035..12051
                     /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(12040..12056,1..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 gcgccrggsg gggsggggsg gggsgrgggg sggggsgggg cgaggcggag aggtgcggcg
     1681 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
     1741 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc
     1801 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
     1861 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
     1921 tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct
     1981 ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg
     2041 tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg
     2101 ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc
     2161 tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg
     2221 ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc
     2281 ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg
     2341 gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggrgggctcg
     2401 ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc
     2461 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt
     2521 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg
     2581 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt
     2641 ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga
     2701 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac
     2761 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg
     2821 tctcatcatt ttggcaaaga attgatttga taccgcgggc tcgacactag tgaacctctt
     2881 cgagggacct aataacttcg tatagcatac attatacgaa gttatattaa gggttccgga
     2941 tccacatgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3001 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3061 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3121 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3181 tctggatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3241 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3301 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3361 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3421 ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3481 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3541 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3601 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     3661 tctgtcgacc tgcagccaag ctagtataac ttcgtatagc atacattata cgaagttatt
     3721 gcggccgcgg actcgccgcc accatggact ataaggacca cgacggagac tacaaggatc
     3781 atgatattga ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg
     3841 gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca
     3901 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg
     3961 tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg
     4021 acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca
     4081 gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg
     4141 acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg
     4201 agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca
     4261 ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga
     4321 tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc
     4381 tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc
     4441 agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg
     4501 ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga
     4561 agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga
     4621 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg
     4681 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca
     4741 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca
     4801 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc
     4861 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc
     4921 agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca
     4981 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga
     5041 acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga
     5101 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga
     5161 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc
     5221 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca
     5281 ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc
     5341 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc
     5401 tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa
     5461 tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca
     5521 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt
     5581 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat
     5641 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg
     5701 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg
     5761 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc
     5821 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc
     5881 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca
     5941 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt
     6001 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta
     6061 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc
     6121 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac
     6181 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc
     6241 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt
     6301 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt
     6361 ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca
     6421 acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag
     6481 aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc
     6541 agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg
     6601 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga
     6661 tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga
     6721 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca
     6781 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg
     6841 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca
     6901 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg
     6961 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca
     7021 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt
     7081 gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata
     7141 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga
     7201 ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct
     7261 tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt
     7321 ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct
     7381 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc
     7441 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc
     7501 tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga
     7561 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc
     7621 agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca
     7681 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct
     7741 acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta
     7801 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga
     7861 agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg
     7921 gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaaaagg ccggcggcca
     7981 cgaaaaaggc cggccaggca aaaaagaaaa agtgataagg cctcgagcct cgactgtgcc
     8041 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     8101 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     8161 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     8221 caatagcagg catgctgggg atgcggtggg ctctatggct tctgagcata ggatccccga
     8281 attcgagtct tctgggcagg cttaaaggct aacctggtgc gtggggcgtt gtcctgcaga
     8341 ggaattgaac aggtgtaaaa ttggaggggc aagacttccc acagattttc gattgtgttg
     8401 ttaagtattg taataggggc aaataaggga aatagactag gcactcacct ggggttttat
     8461 gcagcaaaac tacaggttat tattgcttgt gatccgccct ggagaatttt tcaccgaggt
     8521 agattgaaga catgcccacc caaattttaa tattcttcca cttgcgatcc ttgctacagt
     8581 atgaaattac agtatcgtga attagaatat ataagcagaa ttttaagcat tttaaaagag
     8641 cccagtactt catgtctgtc tctcccactt ctgcagccct atcaaagggt attttagcac
     8701 actcatttta gccccatttt catttgttgt actggcttat ccaatcccta gacagagcac
     8761 tggcattccc tctctcctga tcttagaagt ccgatgactc atgaaaccag acagattagt
     8821 ttcatacacc acaaatcgag gctgtagctg gggcctttaa cattgcagtt tttttattct
     8881 tcagtacact ttgttgattc tttgccttga tcttgacttc aggttctatc accaccccct
     8941 cagatggtgt tccacacttg ggcctattca cagttcagag agctttacaa caatagatgt
     9001 attgagaatc caacctaaag ttcagctttt tactcccatg aatgcctctt tcctttttct
     9061 ccatttataa actgagccat ttcctgttaa tggtttacag atgaatatct cctcccccaa
     9121 tatcacctga tgtatcttac attttgccag gcttagattg tcttaaaagg tacataaatt
     9181 aacatgtgaa atttactcct taatgcttca gtggatttca tgagtgcagt acagaagact
     9241 ggtaatgggc taataacttt tatttcaggt acccaattcg ccctatagtg agtcgtatta
     9301 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     9361 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     9421 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     9481 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     9541 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     9601 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     9661 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     9721 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     9781 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     9841 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     9901 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     9961 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
    10021 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
    10081 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
    10141 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
    10201 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
    10261 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
    10321 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
    10381 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
    10441 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
    10501 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
    10561 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
    10621 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
    10681 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
    10741 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
    10801 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
    10861 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
    10921 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
    10981 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
    11041 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
    11101 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
    11161 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
    11221 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
    11281 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
    11341 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
    11401 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
    11461 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
    11521 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
    11581 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
    11641 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
    11701 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
    11761 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
    11821 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
    11881 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
    11941 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
    12001 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttca
//
