LOCUS pACT1:Cas9-GFP, U6:sgUPRT 9983 bp DNA circular SYN 14-APR-2025
DEFINITION Expresses Cas9 fused with GFP and a sgRNA targeting the
Cryptosporidium parvum UPRT gene.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9983)
AUTHORS Wilke G, Funkhouser-Jones LJ, Wang Y, Ravindran S, Wang Q, Beatty
WL, Baldridge MT, VanDussen KL, Shen B, Kuhlenschmidt MS,
Kuhlenschmidt TB, Witola WH, Stappenbeck TS, Sibley LD
TITLE A Stem-Cell-Derived Platform Enables Complete Cryptosporidium
Development In Vitro and Genetic Tractability.
JOURNAL Cell Host Microbe. 2019 Jun 18. pii: S1931-3128(19)30252-5. doi:
10.1016/j.chom.2019.05.007.
PUBMED 31231046
REFERENCE 2 (bases 1 to 9983)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Host
Microbe. 2019 Jun 18. pii: S1931-3128(19)30252-5. doi:
10.1016/j.chom.2019.05.007."
COMMENT Sequence Label: pACT1:Cas9-GFP, U6:sgUPRT
FEATURES Location/Qualifiers
source 1..9983
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 18..37
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 231..253
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 245..262
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 246..262
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 281..299
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 281..298
/label=SP6
/note="SP6 promoter, forward primer"
misc_RNA complement(339..414)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 2082..6185
/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 2082..6185
/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 6189..6215
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 6195..6221
/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 6234..6254
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6261..6281
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS join(6321..6323,6324..7037)
/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 6971..6992
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(7629..7645)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7629..7645)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7642..7664)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7653..7669
/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(7677..7683,7684..7701,7702..7707))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7722..7743
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7860..7877)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8031..8619)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8111..8130)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(8790..9581,9582..9650))
/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 9413..9432
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9651..9755)
/gene="bla"
/label=AmpR promoter
primer_bind 9823..9841
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(9879..9901)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
61 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
121 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
181 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
241 acgttgtaaa acgacggcca gtgaattcga gcgccaagct atttaggtga cactatagaa
301 tactcaagct atgcatcaag cttcctagga aaaaaaaagc accgactcgg tgccactttt
361 tcaagttgat aacggactag ccttatttta acttgctatt tctagctcta aaacattcta
421 catcctctgc ataccccaac acttaacctt tcagttttta tattaaaaaa aactttattt
481 aacttcataa tgacgtgtct gggtcttggc gcctaaatgt taataataaa ccggtattaa
541 gaaatggttt gcataaaatt tgcatgcaat gacaggaatt gagaattttt gcatgtggaa
601 gcggaataaa tagtaaagga tagaatttta agagtagagt ttgaataaaa aatattattg
661 attgataata atacgaatta aatattaaaa aaagagagtg aataaatgtt tgaatactaa
721 aaaaaatata taagtataaa atcaattaat gctgcaatat aatgagcata taagagatat
781 gggagaatac tatagaggaa tagttccaat agtatatgga agtaaaatat tcccaggaga
841 agagaaaaat cctgaagaag agtatttctt gactatattg ttgtgagaaa tgtaggggag
901 ggagatttgg ttaataattt gcaagagttc attgtgttga atagaagctt gtttgtcttt
961 aaagaatgac tgttgaagac tagaattttt ccattcttca tttctatata attggttgaa
1021 gttggaaaga ataattgaag actggttgga gaaatttggg agctcggtac ccggggatcc
1081 tctagagtcg acctgcagtg ggcttcagaa tgagttggtt ataaacagta ataatagaaa
1141 gcgttagggc tataaaaatg accaaataag tcatttagca tcataagtat atacattaga
1201 tacagaatga gttcaaaata aataaaaaat tagacttact tacaaattct aagctatagt
1261 ataaaagaac taagaagatt tgatgaattt gctataccat tttaaatatt tgtttatagt
1321 tttttgatat taatataagg actattaagt gaaaaggaaa ctacacatat ttccgccaat
1381 tcattgtatt gcatcgcaca cataaatgaa agattaaaag aaaagaaatt ctattatcta
1441 gtattaaaat aataccaata ataagcaaca cttttttgtt attttttatt ataaagatta
1501 atttgctact ttaaattgaa aaaaacttta caggtttata caaattcgta catcacatgt
1561 atacaatgct acatacatta cttgataaat attacgacgt gaaacgtatt aatttaaacg
1621 aatcttagtt atgtaaataa aaaaaggtta gattttagta aaaaaaaaag accaaatttt
1681 atattaaata taaatcttgc attgaatgca tttatgtcaa attgcatgca ccttgccgcc
1741 caccatgcat agacgcgcgc caatacgtat taatagacga ttacatgaca aagtttagag
1801 attaaataaa aatcaagtat aaatttagaa tagacaacat gtaaaaagtg agatgaatct
1861 ttttacgata cgtacagtgc gcattaaaaa tccaaagtat atttttattt tcctgcgtgc
1921 gtatacatat ctttaaatac ttggataaag agataataag aattgtacgg caagcaaagt
1981 agtagtttaa tagagcagtt tttatatttt atatatagct tacttacaat atagattttt
2041 ttttttcata gatttcgggc atattacaca agtaacaaaa gatggacaag aagtacagca
2101 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
2161 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
2221 tgatcggcgc cctgctgttc gacagcggag aaacagccga ggccacccgg ctgaagagaa
2281 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
2341 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
2401 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
2461 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
2521 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
2581 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
2641 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
2701 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
2761 cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg
2821 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
2881 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
2941 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
3001 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
3061 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
3121 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatc gatggcggag
3181 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
3241 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
3301 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
3361 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
3421 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
3481 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgcca
3541 gcgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
3601 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtacaacgag ctgaccaaag
3661 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaaag
3721 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
3781 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
3841 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
3901 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
3961 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
4021 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
4081 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
4141 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
4201 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
4261 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
4321 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
4381 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
4441 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
4501 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
4561 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
4621 tgaaggacga ctccatcgat aacaaagtgc tgactcggag cgacaagaac cggggcaaga
4681 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cgccagctgc
4741 tgaatgccaa gctgattacc cagaggaagt tcgacaatct gaccaaggcc gagagaggcg
4801 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
4861 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaacg
4921 acaaactgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
4981 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
5041 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
5101 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
5161 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
5221 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
5281 gcgaaacagg cgagatcgtg tgggataagg gccgggactt tgccaccgtg cggaaagtgc
5341 tgtctatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
5401 aagagtctat cctgcccaag aggaacagcg acaagctgat cgccagaaag aaggactggg
5461 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
5521 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
5581 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
5641 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
5701 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
5761 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
5821 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaaa cactacctgg
5881 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
5941 tggacaaggt gctgagcgcc tacaacaagc acagagacaa gcctatcaga gagcaggccg
6001 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
6061 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
6121 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
6181 gcgacgccta tccctatgac gtgcccgatt atgccagcct gggcagcggc tcccccaaga
6241 aaaaacgcaa ggtggaagat cctaagaaaa agcggaaagt ggacggcatt ggtagtggga
6301 gcaacggcag cagcggatcc gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
6361 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgcgc ggcgagggcg
6421 agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
6481 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
6541 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
6601 aggagcgcac catctccttc aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt
6661 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
6721 gcaacatcct ggggcacaag ctggagtaca acttcaacag ccacaacgtc tatatcacgg
6781 ccgacaagca gaagaacggc atcaaggcga acttcaagat ccgccacaac gtcgaggacg
6841 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
6901 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga
6961 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
7021 acgagctgta caagtagatc aatattgttc ttggagttgt tcttaacagc ttatttcctg
7081 aacacagata cgtattgcaa cttaaataga aatatgaatt acacaagtta tcacttcttg
7141 tagacaattt cttattacat tattcagatc tcagttctat tctcatttga aagctaaaaa
7201 aaaagatgat aaattgtcta atcaatgcta ataaactcta aatttagcaa gaaaattcta
7261 ttataattgt tagtcaatga taactaatat tatggataga aggtgggcta gtttgaatca
7321 aaatagttac taagtaatgt gtagttcgat acgatttttt ccttcttttt aaacgaaagt
7381 gtttaaatta tgtgcgtgtt gttttcctga gcacagtatt agaaattaat taagttgatt
7441 tgtcaggtga tgtttattac caaagttcaa gtccgaaaat agagctgcca tttaacaata
7501 aaaaagagcg gtccaaattt aggcaaaaca aattttttaa aaaaagctcc aagtccggga
7561 attgaacccg ggtcatccgc gtgacaggcg gatatactat gcaggcatgc aagcttggca
7621 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
7681 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
7741 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
7801 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
7861 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
7921 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
7981 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
8041 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
8101 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
8161 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
8221 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
8281 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
8341 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
8401 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
8461 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
8521 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
8581 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
8641 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
8701 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
8761 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
8821 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
8881 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
8941 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
9001 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
9061 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
9121 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
9181 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
9241 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
9301 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
9361 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
9421 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
9481 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
9541 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
9601 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
9661 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
9721 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
9781 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
9841 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
9901 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
9961 tcagcgggtg ttggcgggtg tcg
//
