LOCUS       sequence-118432-       10115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CAS9 in Leishmania.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10115)
  AUTHORS   Zhang WW, Matlashewski G
  TITLE     CRISPR-Cas9-Mediated Genome Editing in Leishmania donovani.
  JOURNAL   MBio. 2015 Jul 21;6(4). pii: e00861-15. doi: 10.1128/mBio.00861-15.
  PUBMED    26199327
REFERENCE   2  (bases 1 to 10115)
  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: "MBio."; 
            date: "2015-07-21"; volume: "6(4). pii: e00861-15. doi"; pages: " 
            10.1128/mBio.00861-15"
COMMENT     Sequence Label: pLPhygCAS9
FEATURES             Location/Qualifiers
     source          1..10115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..20
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2..19
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1206..1215
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1215..1280
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1287..1307
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1332..5432
                     /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             1332..5432
                     /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             5433..5480
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5508..5525)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             6725..7750
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             6725..7750
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(7770..7789)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7771..7789)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7876..7893)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8047..8635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8127..8146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8806..9597,9598..9666))
                     /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     9429..9448
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9667..9771)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9839..9857
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(9895..9917)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     10017..10036
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gatttaggtg acactataga actcgaccgc tgcttgcgtt cgcatggacg ccacgttcga
       61 cccgatcggg cagcaggccg cggagacgcg gcagcgcaaa cgtgcccggc gggtggctcg
      121 ttccggagga gcgttccgcc gtcgctacat cagcatgcaa gaggatcatt gctttgcggt
      181 tgaggtggcg gagctcaact accagcgtgc cgaggaatac gttcatcaag cgtcagccgc
      241 cggcgcggtc gcgacacctg ctcggcggag cggccactgt gctgctctct tgacggccgg
      301 tcgcgacgca gggccgctgg tgggtctcca gcgggtacgg tgcgatcagg cagccatgac
      361 ggctacgctg agcacgcttg cgaaggagct ggctgcactg tcgccgtagc gcgatgcgag
      421 acgtatcgag cgcatgccag tggggtgctg ggcaacactt ggcggcgcgt ggctctttct
      481 cactggtttg ggcacgccca cgctgtgggt gtgtacctgt ctgccctgcc gctctctctc
      541 ctgatctctc gtcctggcaa caacaacaca ccgcgaatga attgcccaca agccatctgc
      601 agtccgcacg cgcgtccggg ccttgcggtg tggatgcgag catcgcgcat tgccggccac
      661 cgccgcgtgc caaaagcaca cacatcgctc cccctcacgt ggtgagcgtc tgcgcgtgtg
      721 tgtctgtgtg cataaatgcg tacgtgtgcg ctcggtccac ggtcaccatc gcgccccgcg
      781 gcatcggtgc tcgcaccttt ccattctctc cgtacggcgt tcgccactct gctaactctt
      841 ctcccctttc ctctctccgc atctgcgcct tgtgtgccgt gtgtgcgccc cacacgtact
      901 ctacacgcct tctctgcatg cgctcctcgc ctgcctgaaa cgatgacgtg cgccgccact
      961 cgctcacccg ctcccatcca catgcattcc tgcaccctac agcgactcta agccgtacat
     1021 tccatctcct caacacccac tcgcttcctt ctgctctcac ctattacttc gccagcccac
     1081 atatctgcta taacctgccc tcccccaccc gcttcccaca catccgccac cgctacgaag
     1141 cctcgactct agaccatgga agcttagcac ctgcctgaaa tcactttttt tcaggttgga
     1201 ccggtgccac catggactat aaggaccacg acggagacta caaggatcat gatattgatt
     1261 acaaagacga tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag
     1321 tcccagcagc cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct
     1381 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
     1441 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
     1501 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
     1561 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
     1621 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
     1681 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
     1741 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
     1801 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
     1861 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
     1921 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
     1981 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     2041 tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     2101 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     2161 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     2221 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     2281 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     2341 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     2401 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     2461 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     2521 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     2581 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     2641 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     2701 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     2761 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     2821 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     2881 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     2941 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     3001 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     3061 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     3121 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     3181 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     3241 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     3301 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     3361 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     3421 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     3481 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     3541 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     3601 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
     3661 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     3721 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     3781 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     3841 tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     3901 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     3961 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
     4021 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     4081 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     4141 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     4201 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     4261 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     4321 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     4381 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
     4441 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
     4501 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
     4561 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
     4621 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
     4681 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
     4741 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
     4801 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
     4861 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
     4921 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
     4981 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
     5041 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
     5101 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
     5161 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
     5221 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
     5281 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
     5341 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
     5401 cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg
     5461 gccaggcaaa aaagaaaaag taagaattcc tagagctcgc tgatcagcct cgactgtgcc
     5521 ttctagttgc cagccatctg ttggatccgc ggccccgcct cgagcccggg cgccgtcgac
     5581 cgctgcttgc gttcgcatgg acgccacgtt cgacccgatc gggcagcagg ccgcggagac
     5641 gcggcagcgc aaacgtgccc ggcgggtggc tcgttccgga ggaggcgttc cgccgtcgct
     5701 acgtcagcat gcaagaggat cattgctttg cggttgaggt ggcggagctc aactaccagc
     5761 gtgccgtgga atacgttcat caagcgtcag ccgccggcgc ggtcgcgaca cctgctcggc
     5821 ggagcggcca ctgtgctgct ctcttgacgg ccggtcgcga cgcagggccg ctggtgggtc
     5881 tccagcgggt acggtgcgat caggcagcca tgacggctac gctgagcacg cttgcgaagg
     5941 agctggctgc actgtcgccg tagcgcgatg cgagacgtat cgagcgcatg ccagtggggt
     6001 gctgggcaac acttggcggc gcgtggctct ttctcactgg tttgggcacg cccacgctgt
     6061 gggtgtgtac ctgtctgccc tgccgctctc tctcctgatc tctcgtcctg gcaacaacaa
     6121 cacaccgcga atgaattgcc cacaagccat ctgcagtccg cacgcgcgtc cgggccttgc
     6181 ggtgtggatg cgagcatcgc gcattgccgg ccaccgccgc gtgccaaaag cacacacatc
     6241 gctccccctc acgtggtgag cgtctgcgcg tgtgtgtctg tgtgcataaa tgcgtacgtg
     6301 tgcgctcggt ccacggtcac catcgcgccc cgcggcatcg gtgctcgcac ctttccattc
     6361 tctccgtacg gcgttcgcca ctctgctaac tcttctcccc tttcctctct ccgcatctgc
     6421 gccttgtgtg ccgtgtgtgc gccccacacg tactctacac gccttctctg catgcgctcc
     6481 tcgcctgcct gaaacgatga cgtgcgccgc cactcgctca cccgctccca tccacatgca
     6541 ttcctgcacc ctacagcgac tctaagccgt acattccatc tcctcaacac ccactcgctt
     6601 ccttctgctc tcacctatta cttcgccagc ccacatatct gctataacct gccctccccc
     6661 acccgcttcc cacacatccg ccaccgctac gaagcctcgg taccgagctc gaattcgcat
     6721 cgatatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     6781 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     6841 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     6901 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     6961 cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     7021 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     7081 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     7141 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     7201 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     7261 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     7321 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     7381 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     7441 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     7501 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     7561 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     7621 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     7681 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     7741 aaaggaatag atatcagatc tgccggtctc cctatagtga gtcgtattaa tttcgataag
     7801 ccaggttaac ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     7861 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     7921 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7981 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     8041 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     8101 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     8161 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     8221 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     8281 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     8341 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     8401 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     8461 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     8521 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     8581 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     8641 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     8701 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     8761 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     8821 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8881 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8941 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     9001 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     9061 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     9121 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     9181 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     9241 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     9301 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     9361 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     9421 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     9481 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     9541 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     9601 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     9661 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     9721 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     9781 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     9841 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     9901 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     9961 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
    10021 atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag aacgcggcta
    10081 caattaatac ataaccttat gtatcataca catac
//
