LOCUS       sequence-102739-       10063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cas9 in T. cruzi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10063)
  AUTHORS   Peng D, Kurup SP, Yao PY, Minning TA, Tarleton RL
  TITLE     CRISPR-Cas9-Mediated Single-Gene and Gene Family Disruption in 
            Trypanosoma cruzi.
  JOURNAL   MBio. 2014 Dec 30;6(1). pii: e02097-14. doi: 10.1128/mBio.02097-14.
  PUBMED    25550322
REFERENCE   2  (bases 1 to 10063)
  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: "2014-12-30"; volume: "6(1). pii: e02097-14. doi"; pages: " 
            10.1128/mBio.02097-14"
COMMENT     Sequence Label: pTrex-b-NLS-hSpCas9
FEATURES             Location/Qualifiers
     source          1..10063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(33..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1695..1760
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1767..1787
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1812..5912
                     /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             1812..5912
                     /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             5913..5960
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6705..7103
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(7878..7896)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7917..7933)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7917..7933)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7930..7952)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7941..7957
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7965..7971,7972..7989,7990..7995))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8010..8031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8148..8165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8319..8907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8399..8418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9078..9869,9870..9938))
                     /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     9701..9720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9939..10043)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcgg
      661 tgcaccctgg gatatttgcg cacccacctt tcccgtcgca atcggcgccg tccgaacgcg
      721 gaaatgttcc gagaataata cggctgggct gcgccgcgcg ttccccgcgt gaaccaacta
      781 ttgtgcgatg tgcagcatcc tattgtggtg ctttaccgca caccagtttc cttgcggggc
      841 ttatttggtg tgttggattt ttaatttttt gatgattcgg aggaattttt tgcaattttt
      901 gggtacccct aaaaaaattt tggagtttcc ggggttcgat ccaacttgtg cttttttaac
      961 cgtccatttg tgtctagtac atcatggagc ggtattctct ccggtggccc aggtggcgtc
     1021 ggaaggaatc ggtgtccctc aggatgtttg gtttttcgtt tttttttttg cggtcttttg
     1081 tttcgcatcg ccgtatcgtg tgcggcgcga ccttcttttt ggctcattgg tcagtgaggt
     1141 ttataataaa gtggatcatt tgggtttttg gccctttgcg gcgcacccgc ggaatggggg
     1201 agggattttc atataattgt agtgtacaca ttgtgtgtgg gtcgcaaaat gtcatgccgt
     1261 gtgcaatgtt tatttgcgtg ctgtgcgtgt ccttttgtga ttccaccact tgtgtgtatt
     1321 ttaccttatc gtctctccct cgcgttggct gtgcgtgaat tgtctgtcgc attcgtggga
     1381 aatcaaaagg ggaatcaacg tgttcttttt ttttcagaag gggagtcaaa gcgccgtgtg
     1441 gatgccaagt ctgtgcaaga gttctagtgg atcccccaac gagtttcttc aaaatatgca
     1501 gcggattcac taagaaacat tttcacgcac gaaagcgaaa ttattatgat tgttattata
     1561 atactttttc tttgttgttt tatccactta ttattgttgt gttaaattgt ttttaccttt
     1621 tttcttttcc aacttctttt atgatgtctt ttcttttttt tttttttgct ctataagttg
     1681 tcttgtctag aatggactat aaggaccacg acggagacta caaggatcat gatattgatt
     1741 acaaagacga tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag
     1801 tcccagcagc cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct
     1861 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
     1921 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
     1981 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
     2041 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
     2101 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
     2161 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
     2221 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
     2281 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
     2341 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
     2401 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
     2461 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     2521 tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     2581 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     2641 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     2701 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     2761 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     2821 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     2881 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     2941 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     3001 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     3061 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     3121 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     3181 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     3241 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     3301 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     3361 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     3421 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     3481 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     3541 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     3601 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     3661 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     3721 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     3781 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     3841 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     3901 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     3961 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     4021 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     4081 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
     4141 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     4201 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     4261 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     4321 tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     4381 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     4441 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
     4501 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     4561 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     4621 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     4681 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     4741 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     4801 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     4861 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
     4921 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
     4981 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
     5041 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
     5101 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
     5161 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
     5221 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
     5281 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
     5341 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
     5401 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
     5461 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
     5521 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
     5581 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
     5641 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
     5701 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
     5761 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
     5821 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
     5881 cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg
     5941 gccaggcaaa aaagaaaaag taactcgagc catttacgac tccaaggcaa cgctgcagaa
     6001 caacctgccg aaggagcgcc gcttcttcaa gattgtgtcg tggtacgaca acgagtgggg
     6061 atactcccac cgcgtggtgg accttgtacg ccacatggcc tcgaaggatc gttcggcaag
     6121 gttgtaggcg tggcgatgac ttcaggtctt tcttttgcga atagggatct tataatacac
     6181 gatgcgtgtc ccgtgatgat cgttaccggt gctgccacga tccaattgac acagcgtcaa
     6241 gagcaaaaca attttacttt tccctttaag gacaacaaca aaaaaatata taactttttt
     6301 ttcttttttt tttttttttg aaattatatt tatggtcatc tttgggaaac aaaaagcagc
     6361 aatttaatga tgcggaagga tgagtgaaat aatgtttaat caatgtacga ggatttgggg
     6421 tattgcaagg aaaatgtaga tgatttaatt gggtgtgtga tgcagcttgt ggtaattttt
     6481 gctcacttcc ctttttgcca catcttttta gtttttctgc ttttctttcc ccattattcc
     6541 acttgtctct cttttcccac gtttcctgca cgaatgcaga aagtgatatt tttactttga
     6601 aagccatcta ccaacaacaa ttacattgaa cagaatttgg gattgcgaat taattcctgc
     6661 agccccgcgt gtggcctcga acaccgagcg accctgcagc caatatggcc aagcctttgt
     6721 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
     6781 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
     6841 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
     6901 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
     6961 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
     7021 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
     7081 ctggttatgt gtgggagggc taaggggatc gatccggaac aacctgccga aggagcgccg
     7141 cttcttcaag attgtgtcgt ggtacgacaa cgagtgggga tactcccacc gcgtggtgga
     7201 ccttgtacgc cacatggcct cgaaggatcg ttcggcaagg ttgtaggcgt ggcgatgact
     7261 tcaggtcttt cttttgcgaa tagggatctt ataatacacg atgcgtgtcc cgtgatgatc
     7321 gttaccggtg ctgccacgat ccaagtgaca cagtgtcaag agcaaaacaa ttttactttt
     7381 ccctttaagg acaacaacaa aaaaatatat aacttttttt tctttttttt tttttgaaat
     7441 tatatttatg gtcatctttg ggaaacaaaa agcagcaatt taatgatgcg gaaggatgag
     7501 tgaaataatg tttaatcaat gtacgaggat ttggggtatt gcaaggaaaa tgtagatgat
     7561 ttagttgggt gtatgatgca gcttgtggta attttttaca actttgcgtt attggaagat
     7621 tttggttgtt cagtttcttt ttgtttataa ctgactgttt taatttagtg tgattttttt
     7681 tttttaagga tttaagctgt ttttcttttt ttttcttttt cttcactgat ggtgtttaag
     7741 gtgtagttta gtgtttgtgc attatgcgtc gtagtgttgt tgttctttgg actttttcga
     7801 acaatactgg gcattatagc ccacctgtgt atatgccgct agagtatgct agccgtgtgg
     7861 gtacccagct tttgttccct ttagtgaggg ttaatttcga gcttggcgta atcatggtca
     7921 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     7981 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     8041 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     8101 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     8161 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     8221 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     8281 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     8341 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     8401 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     8461 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     8521 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     8581 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     8641 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     8701 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     8761 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8821 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8881 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8941 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     9001 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     9061 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     9121 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     9181 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     9241 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     9301 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     9361 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     9421 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     9481 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     9541 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     9601 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9661 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9721 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9781 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9841 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9901 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9961 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
    10021 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgc
//
