LOCUS       sequence-123989-       11171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fusion gene CAS9-HA-2xNLS-GFP in the pTREX-n backbone.
            This vector is used for cloning a specific sgRNA by BamHI, to be 
            co-expressed with Cas9 for genome editing in Trypanosoma cruzi..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11171)
  AUTHORS   Lander N, Li ZH, Niyogi S, Docampo R
  TITLE     CRISPR/Cas9-Induced Disruption of Paraflagellar Rod Protein 1 and 2 
            Genes in Trypanosoma cruzi Reveals Their Role in Flagellar 
            Attachment.
  JOURNAL   MBio. 2015 Jul 21;6(4). pii: e01012-15. doi: 10.1128/mBio.01012-15.
  PUBMED    26199333
REFERENCE   2  (bases 1 to 11171)
  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: e01012-15. doi"; pages: " 
            10.1128/mBio.01012-15"
COMMENT     Sequence Label: Cas9/pTREX-n
FEATURES             Location/Qualifiers
     source          1..11171
                     /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             1690..5793
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1690..5793
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5797..5823
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5803..5829
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             5842..5862
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5869..5889
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5929..5931,5932..6645)
                     /codon_start=1
                     /product="enhanced GFP (EGFP) derivative that exhibits 
                     robust folding and enhanced photostability (Zhong et al., 
                     2019)"
                     /label=SiriusGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     6579..6600
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6654..6670)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6655..6671)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             7408..8211
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7471..7490)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8081..8100
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     promoter        complement(8986..9004)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9025..9041)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9025..9041)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9038..9060)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9049..9065
                     /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(9073..9079,9080..9097,9098..9103))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9118..9139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9256..9273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9427..10015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9507..9526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10186..10977,10978..11046))
                     /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     10809..10828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11047..11151)
                     /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 gccgttggct gtgcgtgaat tgtgcttcgc 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 tttttgctct ataagttgtc
     1681 ttgtctagaa tggacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc
     1741 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     1801 accgaccggc acagcatcaa gaagaacctg atcggcgccc tgctgttcga cagcggagaa
     1861 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     1921 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     1981 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     2041 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     2101 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     2161 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     2221 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     2281 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     2341 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     2401 ttcggcaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     2461 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     2521 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     2581 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     2641 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     2701 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     2761 ggctacgccg gctacatcga tggcggagcc agccaggaag agttctacaa gttcatcaag
     2821 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     2881 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     2941 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     3001 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     3061 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     3121 ttcgaggaag tggtggacaa gggcgccagc gcccagagct tcatcgagcg gatgaccaac
     3181 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     3241 ttcaccgtgt acaacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     3301 gccttcctga gcggcgagca gaaaaaagcc atcgtggacc tgctgttcaa gaccaaccgg
     3361 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     3421 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     3481 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     3541 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     3601 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     3661 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     3721 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     3781 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     3841 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     3901 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     3961 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     4021 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     4081 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     4141 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     4201 gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgataa caaagtgctg
     4261 actcggagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     4321 aagatgaaga actactggcg ccagctgctg aatgccaagc tgattaccca gaggaagttc
     4381 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     4441 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     4501 cggatgaaca ctaagtacga cgagaacgac aaactgatcc gggaagtgaa agtgatcacc
     4561 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     4621 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     4681 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     4741 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     4801 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     4861 cggaagcggc ctctgatcga gacaaacggc gaaacaggcg agatcgtgtg ggataagggc
     4921 cgggactttg ccaccgtgcg gaaagtgctg tctatgcccc aagtgaatat cgtgaaaaag
     4981 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgac
     5041 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     5101 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     5161 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     5221 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     5281 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     5341 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     5401 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     5461 tttgtggaac agcacaaaca ctacctggac gagatcatcg agcagatcag cgagttctcc
     5521 aagagagtga tcctggccga cgctaatctg gacaaggtgc tgagcgccta caacaagcac
     5581 agagacaagc ctatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     5641 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     5701 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     5761 acacggatcg acctgtctca gctgggaggc gacgcctatc cctatgacgt gcccgattat
     5821 gccagcctgg gcagcggctc ccccaagaaa aaacgcaagg tggaagatcc taagaaaaag
     5881 cggaaagtgg acggcattgg tagtgggagc aacggcagca gcggatctgt gagcaagggc
     5941 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     6001 cacaagttca gcgtgcgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg
     6061 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     6121 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     6181 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tctccttcaa ggacgacggc
     6241 acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     6301 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     6361 ttcaacagcc acaacgtcta tatcacggcc gacaagcaga agaacggcat caaggcgaac
     6421 ttcaagatcc gccacaacgt cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     6481 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     6541 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     6601 accgccgccg ggatcactct cggcatggac gagctgtaca agtagaagct tatcgatacc
     6661 gtcgacctcg agccatttac gactccaagg caacgctgca gaacaacctg ccgaaggagc
     6721 gccgcttctt caagattgtg tcgtggtacg acaacgagtg gggatactcc caccgcgtgg
     6781 tggaccttgt acgccacatg gcctcgaagg atcgttcggc aaggttgtag gcgtggcgat
     6841 gacttcaggt ctttcttttg cgaataggga tcttataata cacgatgcgt gtcccgtgat
     6901 gatcgttacc ggtgctgcca cgatccaatt gacacagcgt caagagcaaa acaattttac
     6961 ttttcccttt aaggacaaca acaaaaaaat atataacttt tttttctttt tttttttttt
     7021 ttgaaattat atttatggtc atctttggga aacaaaaagc agcaatttaa tgatgcggaa
     7081 ggatgagtga aataatgttt aatcaatgta cgaggatttg gggtattgca aggaaaatgt
     7141 agatgattta attgggtgtg tgatgcagct tgtggtaatt tttgctcact tccctttttg
     7201 ccacatcttt ttagtttttc tgcttttctt tccccattat tccacttgtc tctcttttcc
     7261 cacgtttcct gcacgaatgc agaaagtgat atttttactt tgaaagccat ctaccaacaa
     7321 caattacatt gaacagaatt tgggattgcg aattaattcc tgcagccccg cgtgtggcct
     7381 cgaacaccga gcgaccctgc agccaatatg ggatcggcca ttgaacaaga tggattgcac
     7441 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     7501 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     7561 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     7621 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     7681 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     7741 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     7801 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     7861 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     7921 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     7981 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     8041 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     8101 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     8161 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg aggggatcga
     8221 tccggaacaa cctgccgaag gagcgccgct tcttcaagat tgtgtcgtgg tacgacaacg
     8281 agtggggata ctcccaccgc gtggtggacc ttgtacgcca catggcctcg aaggatcgtt
     8341 cggcaaggtt gtaggcgtgg cgatgacttc aggtctttct tttgcgaata gggatcttat
     8401 aatacacgat gcgtgtcccg tgatgatcgt taccggtgct gccacgatcc aagtgacaca
     8461 gtgtcaagag caaaacaatt ttacttttcc ctttaaggac aacaacaaaa aaatatataa
     8521 cttttttttc tttttttttt tttgaaatta tatttatggt catctttggg aaacaaaaag
     8581 cagcaattta atgatgcgga aggatgagtg aaataatgtt taatcaatgt acgaggattt
     8641 ggggtattgc aaggaaaatg tagatgattt agttgggtgt atgatgcagc ttgtggtaat
     8701 tttttacaac tttgcgttat tggaagattt tggttgttca gtttcttttt gtttataact
     8761 gactgtttta atttagtgtg attttttttt tttaaggatt taagctgttt ttcttttttt
     8821 ttctttttct tcactgatgg tgtttaaggt gtagtttagt gtttgtgcat tatgcgtcgt
     8881 agtgttgttg ttctttggac tttttcgaac aatactgggc attatagccc acctgtgtat
     8941 atgccgctag agtatgctag ccgtgtgggt acccagcttt tgttcccttt agtgagggtt
     9001 aatttcgagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     9061 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     9121 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     9181 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     9241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     9301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     9361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     9421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     9481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     9541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     9601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     9661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     9721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     9781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     9841 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     9901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     9961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
    10021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
    10081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
    10141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
    10201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
    10261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
    10321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
    10381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
    10441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
    10501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
    10561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
    10621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
    10681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
    10741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
    10801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
    10861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
    10921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
    10981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
    11041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
    11101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
    11161 ccgaaaagtg c
//
