LOCUS       sequence-256130-       10736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pDEF-SpCas9, Best (-458) Vector for CRISPRi of PyALBA4, PbEF1a 
            promoter, 1st Generation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10736)
  AUTHORS   Walker MP, Lindner SE
  TITLE     Ribozyme-mediated, multiplex CRISPR gene editing and CRISPR 
            interference (CRISPRi) in rodent-infectious Plasmodium yoelii.
  JOURNAL   J Biol Chem. 2019 Jun 14;294(24):9555-9566. doi: 
            10.1074/jbc.RA118.007121. Epub 2019 May 1.
  PUBMED    31043479
REFERENCE   2  (bases 1 to 10736)
  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"; doi: 
            "10.1074/jbc.RA118.007121"; journalName: "J Biol Chem"; date: 
            "2019-06-14- 14"; volume: "294"; issue: "24"; pages: "9555-9566"
COMMENT     Sequence Label: pSL1315
FEATURES             Location/Qualifiers
     source          1..10736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     ncRNA           612..648
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     ncRNA           759..826
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     CDS             1873..1899
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1903..1923
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1933..1956,1957..1959,1960..4515,4516..4518,
                     4519..6033)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(1933..1956,1957..1959,1960..4446,4447..4449,
                     4450..6033)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6037..6057
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     6109..6129
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6698..6717)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6817..6839
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6877..6895)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6963..7067
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7068..7136,7137..7928)
                     /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(7286..7305)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8099..8687
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8588..8607
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8841..8858
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8975..8996
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9011..9016,9017..9034,9035..9041)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     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)."
     primer_bind     9054..9076
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9073..9089
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9073..9089
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggtaccccgc ggtggcggcc gcagcttaat tcttttcgag ctctttatgc ttaagtttac
       61 aatttaatat tcatacttta agtatttttt gtagtatcct agatattgtg ctttaaatgc
      121 tcacccctca aagcaccagt aatattttca tccactgaaa taccattaaa ttttcaaaaa
      181 aatactatgc atataatgtt atacatataa acataaaacg ccatgtaaat caaaaaatat
      241 ataaaaatat gtataaaaat aaatatgcac taaatataag ctaattatgc ataaaaatta
      301 aagtgccctt tattaactag ctagtcgtaa ttatttatat ttctatgtta taaaaaaatc
      361 ctcatataat aatataatta atatatgtaa tgtttttttt attttataat tttaatataa
      421 aataatatgt aaattaattc aaaaaataaa tataattgtt gtgaaacaaa aaacgtaatt
      481 ttttcatttg ccttcaaaat ttaaatttat tttaatattt cctaaaatat atatactttg
      541 tgtataaata tataaaaata tatatttgct tataaataaa taaaaatttt ataaaaatgg
      601 ctagcaatac gctgatgagt ccgtgaggac gaaacgagta agctcgtccg tatttattat
      661 actataacgt ttcagagcta tgctggaaac agcatagcaa gttgaaataa ggctagtccg
      721 ttatcaactt gaaaaagtgg caccgagtcg gtgcttttgg ccggcatggt cccagcctcc
      781 tcgctggcgc cggctgggca acatgcttcg gcatggcgaa tgggacaccg gtgtttttct
      841 tacttatata tttataccaa ttgattgtat ttataactgt aaaaatgtgt atgttgtgtg
      901 catatttttt tttgtgcatg cacatgcatg taaatagcta aaattatgaa cattttattt
      961 tttgttcaga aaaaaaaaac tttacacaca taaaatggct agtatgaata gccatatttt
     1021 atataaatta aatcctatga atttatgacc atattaaaaa tttagatatt tatggaacat
     1081 aatatgtttg aaacaataag acaaaattat tattattatt attattttta ctgttataat
     1141 tatgttgtct cttcaatgat tcataaatag ttggacttga tttttaaaat gtttataata
     1201 tgattagcat agttaaataa aaaaagttga aaaattaaaa aaaaacatat aaacacaaat
     1261 gatgtttttt ccttcaattt ctcgagagct taattctttt cgagctcttt atgcttaagt
     1321 ttacaattta atattcatac tttaagtatt ttttgtagta tcctagatat tgtgctttaa
     1381 atgctcaccc ctcaaagcac cagtaatatt ttcatccact gaaataccat taaattttca
     1441 aaaaaatact atgcatataa tgttatacat ataaacataa aacgccatgt aaatcaaaaa
     1501 atatataaaa atatgtataa aaataaatat gcactaaata taagctaatt atgcataaaa
     1561 attaaagtgc cctttattaa ctagctagtc gtaattattt atatttctat gttataaaaa
     1621 aatcctcata taataatata attaatatat gtaatgtttt ttttatttta taattttaat
     1681 ataaaataat atgtaaatta attcaaaaaa taaatataat tgttgtgaaa caaaaaacgt
     1741 aattttttca tttgccttca aaatttaaat ttattttaat atttcctaaa atatatatac
     1801 tttgtgtata aatatataaa aatatatatt tgcttataaa taaataaaaa ttttataaaa
     1861 atgactagta tgtacccata cgatgttcca gattacgctt cgccgaagaa aaagcgcaag
     1921 gtcgaagcgt ccgacaagaa gtacagcatc ggcctggcca tcggcaccaa ctctgtgggc
     1981 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     2041 accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa
     2101 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     2161 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     2221 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     2281 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     2341 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     2401 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     2461 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     2521 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     2581 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     2641 ttcggcaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     2701 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     2761 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     2821 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     2881 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     2941 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     3001 ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag
     3061 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     3121 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     3181 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     3241 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     3301 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     3361 ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac
     3421 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     3481 ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     3541 gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg
     3601 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     3661 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     3721 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     3781 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     3841 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     3901 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     3961 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     4021 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     4081 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     4141 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     4201 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     4261 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     4321 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     4381 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     4441 gtggacgcca tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg
     4501 accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     4561 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc
     4621 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     4681 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     4741 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc
     4801 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     4861 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     4921 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     4981 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     5041 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     5101 cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc
     5161 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag
     5221 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat
     5281 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     5341 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     5401 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     5461 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     5521 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     5581 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     5641 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     5701 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc
     5761 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac
     5821 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     5881 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     5941 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     6001 acacggatcg acctgtctca gctgggaggc gacagcccca agaagaagag aaaggtggag
     6061 gccagcgggt aggaattcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga
     6121 gtttgtaaca gctgctggga ttacacatgg catggatgaa ctatacaaat aaggatccgt
     6181 ttttcttact tatatattta taccaattga ttgtatttat aactgtaaaa atgtgtatgt
     6241 tgtgtgcata tttttttttg tgcatgcaca tgcatgtaaa tagctaaaat tatgaacatt
     6301 ttattttttg ttcagaaaaa aaaaacttta cacacataaa atggctagta tgaatagcca
     6361 tattttatat aaattaaatc ctatgaattt atgaccatat taaaaattta gatatttatg
     6421 gaacataata tgtttgaaac aataagacaa aattattatt attattatta tttttactgt
     6481 tataattatg ttgtctcttc aatgattcat aaatagttgg acttgatttt taaaatgttt
     6541 ataatatgat tagcatagtt aaataaaaaa agttgaaaaa ttaaaaaaaa acatataaac
     6601 acaaatgatg ttttttcctt caatttcggc gcctgatgcg gtattttctc cttacgcatc
     6661 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     6721 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     6781 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     6841 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     6901 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     6961 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     7021 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     7081 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     7141 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     7201 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     7261 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     7321 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     7381 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     7441 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7501 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7561 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7621 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7681 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7741 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7801 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7861 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7921 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7981 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     8041 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     8101 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     8161 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     8221 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     8281 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     8341 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     8401 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8461 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8521 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8581 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8641 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8701 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8761 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8821 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     8881 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     8941 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     9001 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     9061 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgat aattcctgca
     9121 gcccagctta attcttttcg agctctttat gcttaagttt acaatttaat attcatactt
     9181 taagtatttt ttgtagtatc ctagatattg tgctttaaat gctcacccct caaagcacca
     9241 gtaatatttt catccactga aataccatta aattttcaaa aaaatactat gcatataatg
     9301 ttatacatat aaacataaaa cgccatgtaa atcaaaaaat atataaaaat atgtataaaa
     9361 ataaatatgc actaaatata agctaattat gcataaaaat taaagtgccc tttattaact
     9421 agctagtcgt aattatttat atttctatgt tataaaaaaa tcctcatata ataatataat
     9481 taatatatgt aatgtttttt ttattttata attttaatat aaaataatat gtaaattaat
     9541 tcaaaaaata aatataattg ttgtgaaaca aaaaacgtaa ttttttcatt tgccttcaaa
     9601 atttaaattt attttaatat ttcctaaaat atatatactt tgtgtataaa tatataaaaa
     9661 tatatatttg cttataaata aataaaaaat tttataaaac atagggggat ccatggttgg
     9721 ttcgctaaac tgcatcgtcg ctgtgtccca gaacatgggc atcggcaaga acggggacct
     9781 gccctggcca ccgctcagga acgaatttag atatttccag agaatgacca caacctcttc
     9841 agtagaaggt aaacagaatc tggtgattat gggtaagaag acctggttct ccattcctga
     9901 gaagaatcga cctttaaagg gtagaattaa tttagttctc agcagagaac tcaaggaacc
     9961 tccacaagga gctcattttc tttccagaag tctagatgat gccttaaaac ttactgaaca
    10021 accagaatta gcaaataaag tagacatggt ctggatagtt ggtggcagtt ctgtttataa
    10081 ggaagccatg aatcacccag gccatcttaa actatttgtg acaaggatca tgcaagactt
    10141 tgaaagtgac acgttttttc cagaaattga tttggagaaa tataaacttc tgccagaata
    10201 cccaggtgtt ctctctgatg tccaggagga gaaaggcatt aagtacaaat ttgaagtata
    10261 tgagaagaat gattaaggat cccgtttttc ttacttatat atttatacca attgattgta
    10321 tttataactg taaaaatgtg tatgttgtgt gcatattttt ttttgtgcat gcacatgcat
    10381 gtaaatagct aaaattatga acattttatt ttttgttcag aaaaaaaaaa ctttacacac
    10441 ataaaatggc tagtatgaat agccatattt tatataaatt aaatcctatg aatttatgac
    10501 catattaaaa atttagatat ttatggaaca taatatgttt gaaacaataa gacaaaatta
    10561 ttattattat tattattttt actgttataa ttatgttgtc tcttcaatga ttcataaata
    10621 gttggacttg atttttaaaa tgtttataat atgattagca tagttaaata aaaaaagttg
    10681 aaaaattaaa aaaaaacata taaacacaaa tgatgttttt tccttcaatt tcgatg
//
