LOCUS       sequence-443898-        8966 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Used for expression of sgRNA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8966)
  TITLE     pSpCas9(BB)-2A-Blast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8966)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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            "Unpublished"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-Blast
FEATURES             Location/Qualifiers
     source          1..8966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1952..1999
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2009..2065
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2066..2461
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(2489..2506)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2495..2702
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2711..2851
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2711..2840
                     /label=AAV2 ITR
     rep_origin      2926..3381
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3013..3032)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3223..3244
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3398..3417)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3517..3539
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3577..3595)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3663..3767
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3768..3836,3837..4628)
                     /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(3986..4005)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4799..5387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5288..5307
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5449..5697
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5449..5469
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5620..5639
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        5716..5791
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        5888..6173
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        6175..6451
                     /label=chicken beta-actin promoter
     intron          6452..6679
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      6691..6700
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6700..6765
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6772..6792
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(6817..8966,1..1951)
                     /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             join(6817..8966,1..1951)
                     /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"
ORIGIN
        1 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
       61 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
      121 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
      181 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
      241 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
      301 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
      361 ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
      421 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
      481 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
      541 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
      601 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
      661 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
      721 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
      781 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
      841 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
      901 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
      961 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     1021 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
     1081 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
     1141 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa
     1201 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     1261 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     1321 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     1381 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     1441 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     1501 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     1561 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     1621 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
     1681 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
     1741 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     1801 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     1861 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     1921 acggatcgac ctgtctcagc tgggaggcga caaaaggccg gcggccacga aaaaggccgg
     1981 ccaggcaaaa aagaaaaagg aattcggcgc aacaaacttc tctctgctga aacaagccgg
     2041 agatgtcgaa gagaatcctg gaccggccaa gcctttgtct caagaagaat ccaccctcat
     2101 tgaaagagca acggctacaa tcaacagcat ccccatctct gaggactaca gcgtcgccag
     2161 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg
     2221 gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct
     2281 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg
     2341 ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg
     2401 acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta
     2461 agaattctaa ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2521 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2581 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2641 ggtggggtgg ggcaggacag caagggggag gattgggaag agaatagcag gcatgctggg
     2701 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     2761 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     2821 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     2881 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     2941 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3001 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3061 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3121 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     3181 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3241 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     3301 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3361 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     3421 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3481 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3541 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3601 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3661 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3721 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3781 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3841 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3901 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3961 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4021 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4081 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4141 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4201 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4261 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4321 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4381 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4441 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg
     4501 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4561 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4621 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4681 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4741 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4801 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4861 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4921 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4981 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5041 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5101 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5161 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5221 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5281 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5341 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5401 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtga gggcctattt
     5461 cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt
     5521 aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt
     5581 cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta
     5641 acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccggg
     5701 tcttcgagaa gacctgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     5761 tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt ttagagctag aaatagcaag
     5821 ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt ctgcagacaa atggctctag
     5881 aggtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     5941 gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     6001 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     6061 ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat
     6121 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
     6181 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
     6241 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg gggggcgcgc
     6301 gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc
     6361 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
     6421 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt
     6481 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
     6541 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagctga gcaagaggta
     6601 agggtttaag ggatggttgg ttggtggggt attaatgttt aattacctgg agcacctgcc
     6661 tgaaatcact ttttttcagg ttggaccggt gccaccatgg actataagga ccacgacgga
     6721 gactacaagg atcatgatat tgattacaaa gacgatgacg ataagatggc cccaaagaag
     6781 aagcggaagg tcggtatcca cggagtccca gcagccgaca agaagtacag catcggcctg
     6841 gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     6901 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga
     6961 gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga
     7021 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
     7081 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
     7141 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
     7201 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
     7261 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
     7321 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
     7381 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
     7441 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
     7501 cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc
     7561 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
     7621 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
     7681 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
     7741 accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac
     7801 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
     7861 attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag
     7921 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
     7981 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc
     8041 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt
     8101 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
     8161 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
     8221 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag
     8281 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
     8341 aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac
     8401 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg
     8461 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
     8521 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
     8581 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
     8641 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
     8701 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
     8761 cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc
     8821 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
     8881 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
     8941 aaagcccagg tgtccggcca gggcga
//
