LOCUS       sequence-249688-        8395 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Cas9 D10A Nickase Vector targeting the 5' end of the mouse Srcap 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8395)
  TITLE     Plasmids for generating AID/Degron Cell Lines in Human and Mouse
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8395)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pX335 Mouse 5' Srcap gRNA B
FEATURES             Location/Qualifiers
     source          1..8395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        270..345
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        398..683
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        685..962
                     /label=chicken beta-actin promoter
     intron          963..1190
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1211..1237
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1241..1261
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1271..1294,1295..1297,1298..5371)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1271..5371
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5375..5395
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5432..5449)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5438..5645
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5654..5794
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5654..5783
                     /label=AAV2 ITR
     rep_origin      5869..6324
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5956..5975)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6166..6187
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6341..6360)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6460..6482
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6520..6538)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6606..6710
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6711..6779,6780..7571)
                     /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(6929..6948)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7742..8330
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8231..8250
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ctcccaatcc tacagacacg ttttagagct agaaatagca agttaaaata
      301 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttctagacga
      361 ttttttagtg cgtgcgccaa ttctgactag aggtacccgt tacataactt acggtaaatg
      421 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag
      481 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      541 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      601 cctggcattg tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      661 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      721 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      781 cgatgggggc gggggggggg ggggggcgcg cgccrggsgg ggcggggcgg ggcggggggc
      841 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      901 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      961 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
     1021 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
     1081 ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg
     1141 tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg
     1201 tgccaccatg tacccatacg atgttccaga ttacgcttcg ccgaagaaaa agcgcaaggt
     1261 cgaagcgtcc gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg
     1321 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
     1381 cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac
     1441 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
     1501 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
     1561 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
     1621 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
     1681 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
     1741 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
     1801 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
     1861 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     1921 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     1981 cggcaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     2041 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     2101 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     2161 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     2221 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     2281 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     2341 ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     2401 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     2461 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     2521 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     2581 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     2641 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     2701 cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt
     2761 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     2821 caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     2881 cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa
     2941 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     3001 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     3061 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     3121 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     3181 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     3241 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     3301 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     3361 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     3421 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     3481 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     3541 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     3601 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     3661 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     3721 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     3781 ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
     3841 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     3901 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
     3961 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     4021 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     4081 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
     4141 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     4201 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     4261 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     4321 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     4381 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     4441 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
     4501 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
     4561 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa
     4621 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     4681 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     4741 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     4801 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     4861 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     4921 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     4981 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     5041 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
     5101 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
     5161 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     5221 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     5281 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     5341 acggatcgac ctgtctcagc tgggaggcga cagccccaag aagaagagaa aggtggaggc
     5401 cagctaagaa ttcctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     5461 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     5521 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     5581 gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct
     5641 ggggagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     5701 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     5761 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     5821 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     5881 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5941 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     6001 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6061 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     6121 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6181 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     6241 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6301 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     6361 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     6421 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     6481 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     6541 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     6601 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6661 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6721 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6781 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6841 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6901 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6961 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     7021 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     7081 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     7141 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     7201 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     7261 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     7321 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     7381 cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca
     7441 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7501 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7561 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     7621 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7681 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7741 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7801 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7861 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7921 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7981 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     8041 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     8101 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     8161 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     8221 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     8281 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     8341 acgcggcctt tttacggttc ctggcctttt gctcacatgt aaggtcgggc aggaa
//
