LOCUS       sequence-180069-        8684 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9 from S. pyogenes with 2A-miRFP670, and cloning backbone for 
            sgRNA. Modified Zhang Plasmid #62988.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8684)
  TITLE     pSpCas9(BB)-2A-miRFP670
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8684)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pSpCas9(BB)-2A-miRFP670
FEATURES             Location/Qualifiers
     source          1..8684
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      226..235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             235..300
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             307..327
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             352..4452
                     /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             352..4452
                     /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             4453..4500
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4516..4569
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4570..5517
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     primer_bind     complement(5551..5568)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5557..5764
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5773..5902
                     /label=AAV2 ITR
     primer_bind     complement(5976..5994)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6062..6166
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6167..6235,6236..7027)
                     /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(6385..6404)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7198..7786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7687..7706
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7848..8096
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7848..7868
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8019..8038
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8115..8190
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8287..8572
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
       61 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
      121 gctgagcaag aggtaagggt ttaagggatg gttggttggt ggggtattaa tgtttaatta
      181 cctggagcac ctgcctgaaa tcactttttt tcaggttgga ccggtgccac catggactat
      241 aaggaccacg acggagacta caaggatcat gatattgatt acaaagacga tgacgataag
      301 atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc cgacaagaag
      361 tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag
      421 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
      481 aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg
      541 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
      601 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
      661 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
      721 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
      781 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
      841 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
      901 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
      961 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
     1021 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg
     1081 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
     1141 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     1201 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
     1261 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga
     1321 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
     1381 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac
     1441 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
     1501 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
     1561 ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
     1621 cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
     1681 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
     1741 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
     1801 ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
     1861 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg
     1921 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
     1981 aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
     2041 aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa
     2101 gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag
     2161 gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca
     2221 ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac
     2281 gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg
     2341 aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag
     2401 tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt
     2461 aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt
     2521 gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg
     2581 gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc
     2641 agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc
     2701 gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc
     2761 cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg
     2821 gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag
     2881 agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg
     2941 ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg
     3001 cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag
     3061 agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc
     3121 cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac
     3181 gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc
     3241 gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc
     3301 cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg
     3361 gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag
     3421 agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac
     3481 tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag
     3541 acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg
     3601 aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc
     3661 ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag
     3721 gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg
     3781 gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg
     3841 gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc
     3901 aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc
     3961 gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac
     4021 gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag
     4081 ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac
     4141 tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac
     4201 gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag
     4261 caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc
     4321 aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac
     4381 gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag
     4441 ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag
     4501 gaattcggca gtggagaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat
     4561 cctggcccaa tggtagcagg tcatgcctct ggcagccccg cattcgggac cgcctctcat
     4621 tcgaattgcg aacatgaaga gatccacctc gccggctcga tccagccgca tggcgcgctt
     4681 ctggtcgtca gcgaacatga tcatcgcgtc atccaggcca gcgccaacgc cgcggaattt
     4741 ctgaatctcg gaagcgtact cggcgttccg ctcgccgaga tcgacggcga tctgttgatc
     4801 aagatcctgc cgcatctcga tcccaccgcc gaaggcatgc cggtcgcggt gcgctgccgg
     4861 atcggcaatc cctctacgga gtactgcggt ctgatgcatc ggcctccgga aggcgggctg
     4921 atcatcgaac tcgaacgtgc cggcccgtcg atcgatctgt caggcacgct ggcgccggcg
     4981 ctggagcgga tccgcacggc gggttcactg cgcgcgctgt gcgatgacac cgtgctgctg
     5041 tttcagcagt gcaccggcta cgaccgggtg atggtgtatc gtttcgatga gcaaggccac
     5101 ggcctggtat tctccgagtg ccatgtgcct gggctcgaat cctatttcgg caaccgctat
     5161 ccgtcgtcga ctgtcccgca gatggcgcgg cagctgtacg tgcggcagcg cgtccgcgtg
     5221 ctggtcgacg tcacctatca gccggtgccg ctggagccgc ggctgtcgcc gctgaccggg
     5281 cgcgatctcg acatgtcggg ctgcttcctg cgctcgatgt cgccgtgcca tctgcagttc
     5341 ctgaaggaca tgggcgtgcg cgccaccctg gcggtgtcgc tggtggtcgg cggcaagctg
     5401 tggggcctgg ttgtctgtca ccattatctg ccgcgcttca tccgtttcga gctgcgggcg
     5461 atctgcaaac ggctcgccga aaggatcgcg acgcggatca ccgcgcttga gagctaagct
     5521 agcgaattct aactagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     5581 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     5641 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     5701 ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg
     5761 gggagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     5821 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     5881 cagtgagcga gcgagcgcgc agctgcctgc aatcactaca ttcacatctg atttcagtag
     5941 ttccttacga gagttcctcc gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     6001 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     6061 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6121 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6181 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6241 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6301 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6361 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6421 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6481 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6541 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6601 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6661 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6721 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6781 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6841 tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc
     6901 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6961 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7021 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7081 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7141 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7201 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7261 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7321 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7381 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7441 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7501 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7561 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7621 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7681 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7741 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7801 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtgag ggcctatttc
     7861 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attggaatta
     7921 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc
     7981 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa
     8041 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgggt
     8101 cttcgagaag acctgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
     8161 caacttgaaa aagtggcacc gagtcggtgc ttttttgttt tagagctaga aatagcaagt
     8221 taaaataagg ctagtccgtt tttagcgcgt gcgccaattc tgcagacaaa tggctctaga
     8281 ggtacccgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     8341 cccattgacg tcaatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     8401 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     8461 tattgacgtc aatgacggta aatggcccgc ctggcattgt gcccagtaca tgaccttatg
     8521 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
     8581 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
     8641 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggg
//
