LOCUS       sequence-431289-       10131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone to express gene specific gRNA from the Plasmodium 
            berghei U6 promoter and the Cas9 nuclease for traditional CRISPR 
            editing..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10131)
  AUTHORS   Jonsdottir TK, Paoletta MS, Ishizaki T, Hernandez S, Ivanova M, 
            Curbelo AH, Saiki PA, Selinger M, Das D, Henriksson J, Bushell ESC
  TITLE     A scalable CRISPR-Cas9 gene editing system facilitates CRISPR 
            screens in the malaria parasite Plasmodium berghei
  JOURNAL   bioRxiv 2024.04.20.590404
REFERENCE   2  (bases 1 to 10131)
  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: "bioRxiv 
            2024.04.20.590404"
COMMENT     Sequence Label: pPbU6_hdhfr/yfcu_Cas9
FEATURES             Location/Qualifiers
     source          1..10131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        11..86
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        126..230
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(231..299,300..1091)
                     /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(449..468)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1262..1850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1751..1770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2004..2021
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2138..2159
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2174..2179,2180..2197,2198..2204)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2212..2228
                     /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     2217..2239
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2236..2252
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2236..2252
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4596..4649
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4659..4724
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4731..4751
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4776..8876
                     /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             4776..8876
                     /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             8877..8924
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(9400..9422)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cctcgtctca gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
       61 ttgaaaaagt ggcaccgagt cggtgctttt tttggacgtc aggtggcact tttcggggaa
      121 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      181 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
      241 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
      301 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
      361 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
      421 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
      481 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
      541 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
      601 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
      661 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
      721 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
      781 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
      841 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
      901 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
      961 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     1021 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     1081 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     1141 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     1201 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     1261 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1321 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     1381 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     1441 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     1501 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     1561 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     1621 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     1681 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     1741 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     1801 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     1861 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     1921 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     1981 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     2041 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2101 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     2161 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     2221 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt ggtacccatg
     2281 ccatggtccc cgcggccgct cgaggaattc cttaagttta caatttaata ttcatacttt
     2341 aagtattttt tgtagtatcc tagatattgt gctttaaatg ctcacccctc aaagcaccag
     2401 taatattttc atccactgaa ataccattaa attttcaaaa aaatactatg catataatgt
     2461 tatacatata aacataaaac gccatgtaaa tcaaaaaata tataaaaata tgtataaaaa
     2521 taaatatgca ctaaatataa gctaattatg cataaaaatt aaagtgccct ttattaacta
     2581 gtcgtaatta tttatatttc tatgttataa aaaaatcctc atataataat ataattaata
     2641 tatgtaatgt tttttttatt ttataatttt aatataaaat aatatgtaaa ttaattcaaa
     2701 aaataaatat aattgttgtg aaacaaaaaa cgtaattttt tcatttgcct tcaaaattta
     2761 aatttatttt aatatttcct aaaatatata tactttgtgt ataaatatat aaaaatatat
     2821 atttgcttat aaataaataa aaaattttat aaaacatagg gggatctatg gttggttcgc
     2881 taaactgcat cgtcgctgtg tcccagaaca tgggcatcgg caagaacggg gacctgccct
     2941 ggccaccgct caggaacgaa tttagatatt tccagagaat gaccacaacc tcttcagtag
     3001 aaggtaaaca gaatctggtg attatgggta agaagacctg gttctccatt cctgagaaga
     3061 atcgaccttt aaagggtaga attaatttag ttctcagcag agaactcaag gaacctccac
     3121 aaggagctca ttttctttcc agaagtctag atgatgcctt aaaacttact gaacaaccag
     3181 aattagcaaa taaagtagac atggtctgga tagttggtgg cagttctgtt tataaggaag
     3241 ccatgaatca cccaggccat cttaaactat ttgtgacaag gatcatgcaa gactttgaaa
     3301 gtgacacgtt ttttccagaa attgatttgg agaaatataa acttctgcca gaatacccag
     3361 gtgttctctc tgatgtccag gaggagaaag gcattaagta caaatttgaa gtatatgaga
     3421 agaatgatgc aagcggagga ggtggatctg gtggaggtgg aagtgcaagc gtgacagggg
     3481 gaatggctag caagtgggat cagaagggta tggacattgc ctatgaggag gcggccttag
     3541 gttacaagga gggtggtgtt cctattggcg gatgtcttat caataacaaa gacggaagtg
     3601 ttctcggtcg tggtcacaac atgagatttc aaaagggatc cgccacacta catggtgaga
     3661 tctccacttt ggaaaactgt gggagattag agggcaaagt gtacaaagat accactttgt
     3721 atacgacgct gtctccatgc gacatgtgta caggtgccat catcatgtat ggtattccac
     3781 gctgtgttgt cggtgagaac gttaatttca aaagtaaggg cgagaaatat ttacaaacta
     3841 gaggtcacga ggttgttgtt gttgacgatg agaggtgtaa aaagatcatg aaacaattta
     3901 tcgatgaaag acctcaggat tggtttgaag atattggtga ggcttcggaa ccatttaaga
     3961 acgtctactt gctacctcaa acaaaccaat tgctgggttt gtacaccatc atcagaaata
     4021 agaatacaac tagacctgat ttcattttct actccgatag aatcatcaga ttgttggttg
     4081 aagaaggttt gaaccatcta cctgtgcaaa agcaaattgt ggaaactgac accaacgaaa
     4141 acttcgaagg tgtctcattc atgggtaaaa tctgtggtgt ttccattgtc agagctggtg
     4201 aatcgatgga gcaaggatta agagactgtt gtaggtctgt gcgtatcggt aaaattttaa
     4261 ttcaaaggga cgaggagact gctttaccaa agttattcta cgaaaaatta ccagaggata
     4321 tatctgaaag gtatgtcttc ctattagacc caatgctggc caccggtggt agtgctatca
     4381 tggctacaga agtcttgatt aagagaggtg ttaagccaga gagaatttac ttcttaaacc
     4441 taatctgtag taaggaaggg attgaaaaat accatgccgc cttcccagag gtcagaattg
     4501 ttactggtgc cctcgacaga ggtctagatg aaaacaagta tctagttcca gggttgggtg
     4561 actttggtga cagatactac tgtgttggtt cgggagaggg cagaggatcc ctgctaacat
     4621 gcggtgatgt cgaggagaat cctggcccag aatcgatgga ctataaggac cacgacggag
     4681 actacaagga tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga
     4741 agcggaaggt cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg
     4801 acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
     4861 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
     4921 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
     4981 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     5041 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     5101 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     5161 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     5221 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     5281 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     5341 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     5401 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     5461 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     5521 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     5581 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     5641 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     5701 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     5761 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     5821 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     5881 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     5941 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     6001 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     6061 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     6121 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     6181 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
     6241 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     6301 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
     6361 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
     6421 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     6481 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     6541 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     6601 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     6661 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     6721 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     6781 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     6841 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     6901 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     6961 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     7021 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     7081 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     7141 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
     7201 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
     7261 tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg
     7321 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
     7381 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
     7441 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
     7501 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
     7561 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
     7621 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
     7681 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
     7741 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
     7801 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
     7861 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
     7921 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
     7981 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
     8041 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
     8101 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
     8161 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
     8221 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
     8281 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
     8341 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
     8401 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
     8461 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
     8521 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
     8581 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
     8641 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
     8701 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
     8761 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
     8821 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa
     8881 ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaagtaagaa ttaccggtga
     8941 tcccgttttt cttacttata tatttatacc aattgattgt atttataact gtaaaaatgt
     9001 gtatgttgtg tgcatatttt tttttgtgca tgcacatgca tgtaaatagc taaaattatg
     9061 aacattttat tttttgttca gaaaaaaaaa actttacaca cataaaatgg ctagtatgaa
     9121 tagccatatt ttatataaat taaatcctat gaatttatga ccatattaaa aatttagata
     9181 tttatggaac ataatatgtt tgaaacaata agacaaaatt attattatta ttattatttt
     9241 tactgttata attatgttgt ctcttcaatg attcataaat agttggactt gatttttaaa
     9301 atgtttataa tatgattagc atagttaaat aaaaaaagtt gaaaaattaa aaaaaaacat
     9361 ataaacacaa atgatgtttt ttccttcaat ttcggccgtc gttttacaac gtcgtgactg
     9421 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     9481 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     9541 cgaatgatcg agaactattg tttccttttt tgttttttta tttttaatta aaatgtaaat
     9601 ataaactcac aaaatgaatg aataaataaa taaataaata aataaaaaat gttaaatcgt
     9661 gttaaaccgc gtgcttcatg tgatatggca agcgattaat ggccatttta tccattttta
     9721 taatttgata aaaaattaaa acaaataaat gtaatgttaa ttatattttg tactataaat
     9781 attattctct acttcacgca aatgtaacac tatccaacta cactcataat gtttaaagaa
     9841 aattggcatt gttttaccat ttttttaaaa caaaaaaaga ataaaatact ttcgtattat
     9901 ttagctaaaa ttgtactaaa tttcatacca aaaaaaaata tatatatgta tgcagctaat
     9961 atttatttga taagttttta tgtaaataat atttaattaa aaaaaaaatt acatagcatg
    10021 ccgaatgcac tattcatttt atgggggtaa ttttttgagt aaccattttt actttttttt
    10081 tacatatgcg catacttcga gttatacaat attattggag acgcgtagag g
//
