LOCUS       sequence-201822-        8508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 and Rosa26 locus specific sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8508)
  AUTHORS   Lusk SJ, McKinney A, Hunt PJ, Fahey PG, Patel J, Chang A, Sun JJ, 
            Martinez VK, Zhu PJ, Egbert JR, Allen G, Jiang X, Arenkiel BR, 
            Tolias AS, Costa-Mattioli M, Ray RS
  TITLE     A CRISPR toolbox for generating intersectional genetic mouse models 
            for functional, molecular, and anatomical circuit mapping.
  JOURNAL   BMC Biol. 2022 Jan 28;20(1):28. doi: 10.1186/s12915-022-01227-0.
  PUBMED    35086530
REFERENCE   2  (bases 1 to 8508)
  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: "BMC Biol.";
            date: "2022-01-28"; volume: "20(1):28. doi"; pages: " 
            10.1186/s12915-022-01227-0"
COMMENT     Sequence Label: px330_Rosa_sgRNA
FEATURES             Location/Qualifiers
     source          1..8508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        667..915
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     667..687
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     838..857
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        937..1012
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        1109..1394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1396..1673
                     /label=chicken beta-actin promoter
     intron          1674..1901
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1913..1922
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1922..1987
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1994..2014
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2039..6139
                     /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             2039..6139
                     /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             6140..6187
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(6215..6232)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6221..6428
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6437..6577
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6437..6566
                     /label=AAV2 ITR
     rep_origin      6652..7107
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6739..6758)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6949..6970
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7124..7143)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7243..7265
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7303..7321)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7389..7493
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7494..7562,7563..8354)
                     /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(7712..7731)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      661 acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
      721 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
      781 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
      841 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
      901 aaaggacgaa acaccgactg gagttgcaga tcacgagttt tagagctaga aatagcaagt
      961 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttgt
     1021 tttagagcta gaaatagcaa gttaaaataa ggctagtccg tttttagcgc gtgcgccaat
     1081 tctgcagaca aatggctcta gaggtacccg ttacataact tacggtaaat ggcccgcctg
     1141 gctgaccgcc caacgacccc cgcccattga cgtcaatagt aacgccaata gggactttcc
     1201 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     1261 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     1321 gtgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     1381 tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc
     1441 cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg
     1501 cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg cggggcgggg
     1561 cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta
     1621 tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg
     1681 ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc
     1741 tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct
     1801 gtaattagct gagcaagagg taagggttta agggatggtt ggttggtggg gtattaatgt
     1861 ttaattacct ggagcacctg cctgaaatca ctttttttca ggttggaccg gtgccaccat
     1921 ggactataag gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga
     1981 cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga
     2041 caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac
     2101 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag
     2161 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac
     2221 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct
     2281 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga
     2341 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat
     2401 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact
     2461 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat
     2521 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga
     2581 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa
     2641 cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct
     2701 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat
     2761 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc
     2821 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat
     2881 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct
     2941 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat
     3001 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca
     3061 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta
     3121 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa
     3181 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca
     3241 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat
     3301 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa
     3361 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt
     3421 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt
     3481 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct
     3541 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa
     3601 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg
     3661 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa
     3721 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg
     3781 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa
     3841 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac
     3901 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct
     3961 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct
     4021 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt
     4081 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct
     4141 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga
     4201 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa
     4261 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga
     4321 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa
     4381 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga
     4441 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat
     4501 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt
     4561 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa
     4621 gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta
     4681 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa
     4741 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt
     4801 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa
     4861 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct
     4921 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca
     4981 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc
     5041 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat
     5101 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat
     5161 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct
     5221 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac
     5281 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac
     5341 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag
     5401 aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc
     5461 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga
     5521 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct
     5581 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc
     5641 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa
     5701 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta
     5761 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca
     5821 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct
     5881 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat
     5941 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc
     6001 cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt
     6061 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct
     6121 gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa
     6181 gaaaaagtaa gaattcctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     6241 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     6301 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     6361 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagagaa tagcaggcat
     6421 gctggggagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     6481 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     6541 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     6601 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     6661 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     6721 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     6781 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     6841 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     6901 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     6961 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     7021 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     7081 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     7141 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     7201 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     7261 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     7321 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     7381 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     7441 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     7501 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     7561 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     7621 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     7681 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     7741 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     7801 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     7861 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     7921 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     7981 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     8041 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     8101 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     8161 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta
     8221 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     8281 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     8341 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     8401 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     8461 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaa
//
