LOCUS       sequence-490580-        8508 bp    DNA     circular SYN 24-OCT-2025
DEFINITION  A human codon-optimized SpCas9 and chimeric guide RNA expression 
            plasmid encoding guide for G3BP1 Nterm..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8508)
  TITLE     Bartel lab plasmids 2025
REFERENCE   2  (bases 1 to 8508)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: DBJS-p2.11
FEATURES             Location/Qualifiers
     source          1..8508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..30
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(249..353)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     421..439
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(477..499)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     599..618
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(635..1090)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(772..793)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     984..1003
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1165..1305
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1176..1305
                     /label=AAV2 ITR
     polyA_signal    1314..1521
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1510..1527
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1555..1602)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(1603..5703)
                     /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             complement(1603..5703)
                     /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             complement(5728..5748)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(5755..5820)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      5820..5829
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          5841..6068
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     promoter        complement(6069..6346)
                     /label=chicken beta-actin promoter
     enhancer        6348..6633
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(6730..6805)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(6827..7075)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(6885..6904)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(7055..7075)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(7137..7725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7217..7236)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7896..8508,1..179,180..248))
                     /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"
ORIGIN
        1 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
       61 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
      121 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
      181 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
      241 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      301 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      361 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
      421 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
      481 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
      541 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
      601 gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa tattttgtta
      661 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaatagac cgaaatcggc
      721 aaaatccctt ataaatcaaa agaatagccc gagatagagt tgagtgttgt tccagtttgg
      781 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
      841 cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc
      901 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
      961 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg
     1021 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     1081 cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac agatgcgtaa
     1141 ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg
     1201 ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
     1261 gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc cgctccccag
     1321 catgcctgct attctcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca
     1381 gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga
     1441 cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg
     1501 gctggcaact agaaggcaca gtcgaggctg atcagcgagc tctaggaatt cttacttttt
     1561 cttttttgcc tggccggcct ttttcgtggc cgccggcctt ttgtcgcctc ccagctgaga
     1621 caggtcgatc cgtgtctcgt acaggccggt gatgctctgg tggatcaggg tggcgtccag
     1681 cacctctttg gtgctggtgt acctcttccg gtcgatggtg gtgtcaaagt acttgaaggc
     1741 ggcaggggct cccagattgg tcagggtaaa caggtggatg atattctcgg cctgctctct
     1801 gatgggctta tcccggtgct tgttgtaggc ggacagcact ttgtccagat tagcgtcggc
     1861 caggatcact ctcttggaga actcgctgat ctgctcgatg atctcgtcca ggtagtgctt
     1921 gtgctgttcc acaaacagct gtttctgctc attatcctcg ggggagccct tcagcttctc
     1981 atagtggctg gccaggtaca ggaagttcac atatttggag ggcagggcca gttcgtttcc
     2041 cttctgcagt tcgccggcag aggccagcat tctcttccgg ccgttttcca gctcgaacag
     2101 ggagtactta ggcagcttga tgatcaggtc ctttttcact tctttgtagc ccttggcttc
     2161 cagaaagtcg atgggattct tctcgaagct gcttctttcc atgatggtga tccccagcag
     2221 ctctttcaca ctcttcagtt tcttggactt gcccttttcc actttggcca ccaccagcac
     2281 agaataggcc acggtggggc tgtcgaagcc gccgtacttc ttagggtccc agtccttctt
     2341 tctggcgatc agcttatcgc tgttcctctt gggcaggata gactctttgc tgaagccgcc
     2401 tgtctgcacc tcggtctttt tcacgatatt cacttggggc atgctcagca ctttccgcac
     2461 ggtggcaaaa tcccggccct tatcccacac gatctccccg gtttcgccgt ttgtctcgat
     2521 cagaggccgc ttccggatct cgccgttggc cagggtaatc tcggtcttga aaaagttcat
     2581 gatgttgctg tagaagaagt acttggcggt agccttgccg atttcctgct cgctcttggc
     2641 gatcatcttc cgcacgtcgt acaccttgta gtcgccgtac acgaactcgc tttccagctt
     2701 agggtacttt ttgatcaggg cggttcccac gacggcgttc aggtaggcgt cgtgggcgtg
     2761 gtggtagttg ttgatctcgc gcactttgta aaactggaaa tccttccgga aatcggacac
     2821 cagcttggac ttcagggtga tcactttcac ttcccggatc agcttgtcat tctcgtcgta
     2881 cttagtgttc atccgggagt ccaggatctg tgccacgtgc tttgtgatct gccgggtttc
     2941 caccagctgt ctcttgatga agccggcctt atccagttcg ctcaggccgc ctctctcggc
     3001 cttggtcaga ttgtcgaact ttctctgggt aatcagcttg gcgttcagca gctgccgcca
     3061 gtagttcttc atcttcttca cgacctcttc ggagggcacg ttgtcgctct tgccccggtt
     3121 cttgtcgctt ctggtcagca ccttgttgtc gatggagtcg tccttcagaa agctctgagg
     3181 cacgatatgg tccacatcgt agtcggacag ccggttgatg tccagttcct ggtccacgta
     3241 catatcccgc ccattctgca ggtagtacag gtacagcttc tcgttctgca gctgggtgtt
     3301 ttccacgggg tgttctttca ggatctggct gcccagctct ttgatgccct cttcgatccg
     3361 cttcattctc tcgcggctgt tcttctgtcc cttctgggtg gtctggttct ctctggccat
     3421 ttcgatcacg atgttctcgg gcttgtgccg gcccatcact ttcacgagct cgtccaccac
     3481 cttcactgtc tgcaggatgc ccttcttaat ggcggggctg ccggccagat tggcaatgtg
     3541 ctcgtgcagg ctatcgccct ggccggacac ctgggctttc tggatgtcct ctttaaaggt
     3601 caggctgtcg tcgtggatca gctgcatgaa gtttctgttg gcgaagccgt cggacttcag
     3661 gaaatccagg attgtcttgc cggactgctt gtcccggatg ccgttgatca gcttccggct
     3721 cagcctgccc cagccggtgt atctccgccg cttcagctgc ttcatcactt tgtcgtcgaa
     3781 caggtgggca taggttttca gccgttcctc gatcatctct ctgtcctcaa acagtgtcag
     3841 ggtcagcacg atatcttcca gaatgtcctc gttttcctca ttgtccagga agtccttgtc
     3901 cttgataatt ttcagcagat cgtggtatgt gcccagggag gcgttgaacc gatcttccac
     3961 gccggagatt tccacggagt cgaagcactc gattttcttg aagtagtcct ctttcagctg
     4021 cttcacggtc actttccggt tggtcttgaa cagcaggtcc acgatggcct ttttctgctc
     4081 gccgctcagg aaggcgggct ttctcattcc ctcggtcacg tatttcactt tggtcagctc
     4141 gttatacacg gtgaagtact cgtacagcag gctgtgcttg ggcagcacct tctcgttggg
     4201 caggttctta tcgaagttgg tcatccgctc gatgaagctc tgggcggaag cgcccttgtc
     4261 caccacttcc tcgaagttcc agggggtgat ggtttcctcg ctctttctgg tcatccaggc
     4321 gaatctgctg tttcccctgg ccagagggcc cacgtagtag gggatgcgga aggtcaggat
     4381 cttctcgatc ttttcccggt tgtccttcag gaatgggtaa aaatcttcct gccgccgcag
     4441 aatggcgtgc agctctccca ggtggatctg gtgggggatg ctgccgttgt cgaaggtccg
     4501 ctgcttccgc agcaggtcct ctctgttcag cttcacgagc agttcctcgg tgccgtccat
     4561 cttttccagg atgggcttga tgaacttgta gaactcttcc tggctggctc cgccgtcaat
     4621 gtagccggcg tagccgttct tgctctggtc gaagaaaatc tctttgtact tctcaggcag
     4681 ctgctgccgc acgagagctt tcagcagggt caggtcctgg tggtgctcgt cgtatctctt
     4741 gatcatagag gcgctcaggg gggccttggt gatctcggtg ttcactctca ggatgtcgct
     4801 cagcaggatg gcgtcggaca ggttcttggc ggccagaaac aggtcggcgt actggtcgcc
     4861 gatctgggcc agcaggttgt ccaggtcgtc gtcgtaggtg tccttgctca gctgcagttt
     4921 ggcatcctcg gccaggtcga agttgctctt gaagttgggg gtcaggccca ggctcagggc
     4981 aatcaggttt ccgaacaggc cattcttctt ctcgccgggc agctgggcga tcagattttc
     5041 cagccgtctg ctcttgctca gtctggcaga caggatggcc ttggcgtcca cgccgctggc
     5101 gttgatgggg ttttcctcga acagctggtt gtaggtctgc accagctgga tgaacagctt
     5161 gtccacgtcg ctgttgtcgg ggttcaggtc gccctcgatc aggaagtggc cccggaactt
     5221 gatcatgtgg gccagggcca gatagatcag ccgcaggtcg gccttgtcgg tgctgtccac
     5281 cagtttcttt ctcaggtggt agatggtggg gtacttctcg tggtaggcca cctcgtccac
     5341 gatgttgccg aagatggggt gccgctcgtg cttcttatcc tcttccacca ggaaggactc
     5401 ttccagtctg tggaagaagc tgtcgtccac cttggccatc tcgttgctga agatctcttg
     5461 cagatagcag atccggttct tccgtctggt gtatcttctt ctggcggttc tcttcagccg
     5521 ggtggcctcg gctgtttcgc cgctgtcgaa cagcagggct ccgatcaggt tcttcttgat
     5581 gctgtgccgg tcggtgttgc ccagcacctt gaatttcttg ctgggcacct tgtactcgtc
     5641 ggtgatcacg gcccagccca cagagttggt gccgatgtcc aggccgatgc tgtacttctt
     5701 gtcggctgct gggactccgt ggataccgac cttccgcttc ttctttgggg ccatcttatc
     5761 gtcatcgtct ttgtaatcaa tatcatgatc cttgtagtct ccgtcgtggt ccttatagtc
     5821 catggtggca ccggtccaac ctgaaaaaaa gtgatttcag gcaggtgctc caggtaatta
     5881 aacattaata ccccaccaac caaccatccc ttaaaccctt acctcttgct cagctaatta
     5941 cagcccggag gagaagggcc gtcccgcccg ctcacctgtg ggagtaacgc ggtcagtcag
     6001 agccggggcg ggcggcgcga ggcggcggcg gagcggggca cggggcgaag gcagcgcgca
     6061 gcgactcccg cccgccgcgc gcttcgcttt ttatagggcc gccgccgccg ccgcctcgcc
     6121 ataaaaggaa actttcggag cgcgccgctc tgattggctg ccgccgcacc tctccgcctc
     6181 gccccgcccc gcccctcgcc ccgccccgcc ccgcctggcg cgcgcccccc cccccccccc
     6241 cccccccccc ctgcccaaaa aaattaaaaa ataaaaaaat acaaaattgg gggtggggag
     6301 gggggggaga tggggagagt gaagcagaac gtggggctca cctcgaccat ggtaatagcg
     6361 atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca
     6421 caatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga
     6481 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     6541 tggaaagtcc ctattggcgt tactattgac gtcaatgggc gggggtcgtt gggcggtcag
     6601 ccaggcgggc catttaccgt aagttatgta acgggtacct ctagagccat ttgtctgcag
     6661 aattggcgca cgcgctaaaa acggactagc cttattttaa cttgctattt ctagctctaa
     6721 aacaaaaaag caccgactcg gtgccacttt ttcaagttga taacggacta gccttatttt
     6781 aacttgctat ttctagctct aaaactcacc attgctttgg tcaatcggtg tttcgtcctt
     6841 tccacaagat atataaagcc aagaaatcga aatactttca agttacggta agcatatgat
     6901 agtccatttt aaaacataat tttaaaactg caaactaccc aagaaattat tactttctac
     6961 gtcacgtatt ttgtactaat atctttgtgt ttacagtcaa attaattcca attatctctc
     7021 taacagcctt gtatcgtata tgcaaatatg aaggaatcat gggaaatagg ccctcacatg
     7081 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7141 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7201 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7261 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7321 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7381 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7441 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7501 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7561 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7621 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7681 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7741 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7801 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7861 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7921 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7981 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcggctt
     8041 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8101 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8161 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     8221 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8281 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8341 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8401 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8461 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcg
//
