LOCUS       sequence-111878-        9968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for Cas9 mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9968)
  AUTHORS   Harms DW, Quadros RM, Seruggia D, Ohtsuka M, Takahashi G, Montoliu 
            L, Gurumurthy CB
  TITLE     Mouse Genome Editing Using the CRISPR/Cas System.
  JOURNAL   Curr Protoc Hum Genet. 2014 Oct 1;83:15.7.1-15.7.27. doi: 
            10.1002/0471142905.hg1507s83.
  PUBMED    25271839
REFERENCE   2  (bases 1 to 9968)
  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: "Curr Protoc
            Hum Genet."; date: "2014-10-1"; volume: "83:15.7.1-15.7.27. doi"; 
            pages: " 10.1002/0471142905.hg1507s83"
COMMENT     Sequence Label: pBGK
FEATURES             Location/Qualifiers
     source          1..9968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      956..965
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             965..1030
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1037..1057
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1082..5182
                     /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             1082..5182
                     /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             5183..5230
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5471..5500
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5516..5533
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5556..5573)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5562..5786
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5832..6260
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5919..5938)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6129..6150
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6269..6289)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6274..6604
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6455..6590
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6517..6536
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6671..7465
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6725..6744)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7335..7354
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7641..7762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7678..7697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7732..7751
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7811..7827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7811..7827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7824..7846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7835..7851
                     /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)."
     promoter        complement(join(7859..7865,7866..7883,7884..7889))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7904..7925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8042..8059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8213..8801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8293..8312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8972..9763,9764..9832))
                     /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     9595..9614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9833..9937)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattgca ccggtgccac
      961 catggactat aaggaccacg acggagacta caaggatcat gatattgatt acaaagacga
     1021 tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc
     1081 cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat
     1141 caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca
     1201 cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc
     1261 cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta
     1321 tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct tccacagact
     1381 ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa
     1441 catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa
     1501 actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat
     1561 gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt
     1621 ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat
     1681 caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg
     1741 gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct
     1801 gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga
     1861 tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca
     1921 gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct
     1981 gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat
     2041 gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca
     2101 gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg
     2161 ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga
     2221 aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa
     2281 gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc
     2341 cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga
     2401 gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag
     2461 attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt
     2521 ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa
     2581 cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta
     2641 taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag
     2701 cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt
     2761 gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc
     2821 cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat
     2881 caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct
     2941 gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca
     3001 cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag
     3061 gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga
     3121 tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag
     3181 cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca
     3241 cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt
     3301 gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat
     3361 cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat
     3421 gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt
     3481 ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga
     3541 tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat
     3601 cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga
     3661 caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa
     3721 ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac
     3781 caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct
     3841 ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac
     3901 taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa
     3961 gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta
     4021 ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta
     4081 ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat
     4141 gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa
     4201 catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc
     4261 tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc
     4321 caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca
     4381 gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc
     4441 cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta
     4501 ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa
     4561 agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt
     4621 tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta
     4681 ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca
     4741 gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca
     4801 ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca
     4861 gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat
     4921 cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc
     4981 catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc
     5041 tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga
     5101 ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga
     5161 cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa
     5221 aaagaaaaag taagaattct gcagatatcc agcacagtgg cggccgctcg actatgggca
     5281 ccaaagaacc tgtaaacgtt atctttttaa attgaatgtg cacaaataaa agtttggaaa
     5341 agaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     5401 aaaaaaaaaa aaaaaaaaaa aaaaatacgc atcatacgct gtggcgtacc tcgagtctag
     5461 agggcccttc gaacaaaaac tcatctcaga agaggatctg aatatgcata ccggtcatca
     5521 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     5581 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     5641 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     5701 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     5761 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     5821 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5881 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5941 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     6001 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6061 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6121 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6181 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     6241 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     6301 ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     6361 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6421 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6481 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6541 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6601 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     6661 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     6721 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     6781 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     6841 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     6901 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     6961 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     7021 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     7081 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     7141 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     7201 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     7261 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     7321 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     7381 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     7441 tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag
     7501 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     7561 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     7621 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     7681 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     7741 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     7801 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     7861 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     7921 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     7981 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     8041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     8101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     8161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     8221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     8281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     8341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     8401 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     8461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8641 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8821 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8881 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8941 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     9001 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     9061 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     9121 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     9181 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     9241 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     9301 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     9361 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     9421 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     9481 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     9541 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     9601 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     9661 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     9721 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9781 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9841 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     9901 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     9961 ctgacgtc
//
