| 质粒名称: | pX330-BbsI-PITCh |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 127875 |
| 存储实验室: | Peter Kaiser |
|
载体骨架基本信息 |
|
| 载体名称: | pX3301-2 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 8951bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | 无 |
|
文献引用方法 |
|
| 材料和方法部分: | pX330-BbsI-PITCh was a gift from Peter Kaiser (Addgene plasmid # 127875 ; http://n2t.net/addgene:127875 ; RRID:Addgene_127875) |
| 参考文献部分: |
Microhomology based CRISPR tagging tools for protein tracking, purification, and depletion. Lin DW, Chung BP, Huang JW, Wang X, Huang L, Kaiser P. J Biol Chem. 2019 May 28. pii: RA119.008422. doi: 10.1074/jbc.RA119.008422. 10.1074/jbc.RA119.008422 PubMed 31138654
|
买家导航
pX330-BbsI-PITCh载体质粒基本信息
pX330-BbsI-PITCh质粒图谱载体图谱和pX330-BbsI-PITCh载体序列质粒序列多克隆位点信息
pX330-BbsI-PITCh质粒载体简介
pX330-BbsI-PITCh质粒序列载体序列
LOCUS pX330-BbsI-PITCh 8951 bp DNA circular SYN 25-AUG-2024
DEFINITION For user to clone their gRNA oligos to BbsI site. PITCh gRNA
expression cassette is already build in so there is no need to
perform Golden Gate assembly.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8951)
AUTHORS Lin DW, Chung BP, Huang JW, Wang X, Huang L, Kaiser P
TITLE Microhomology based CRISPR tagging tools for protein tracking,
purification, and depletion.
JOURNAL J Biol Chem. 2019 May 28. pii: RA119.008422. doi:
10.1074/jbc.RA119.008422.
PUBMED 31138654
REFERENCE 2 (bases 1 to 8951)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
Chem. 2019 May 28. pii: RA119.008422. doi:
10.1074/jbc.RA119.008422."
FEATURES Location/Qualifiers
source 1..8951
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 51..336
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
intron 620..847
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 859..868
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 868..933
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 940..960
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 985..5085
/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 985..5085
/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 5086..5133
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
primer_bind complement(5161..5178)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5167..5374
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 5383..5523
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 5383..5512
/label=AAV2 ITR
rep_origin 5598..6053
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5685..5704)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5895..5916
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6070..6089)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6189..6211
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6249..6267)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6335..6439
/gene="bla"
/label=AmpR promoter
CDS 6440..7300
/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(6658..6677)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7471..8059
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7960..7979
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 8121..8369
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
misc_RNA 8388..8463
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 8561..8809
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
misc_RNA 8830..8905
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
ORIGIN
1 cgtttttagc gcgtgcgcca attctgcaga caaatggctc tagaggtacc cgttacataa
61 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
121 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
181 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
241 ggtaaatggc ccgcctggca ttgtgcccag tacatgacct tatgggactt tcctacttgg
301 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
361 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
421 ttattttgtg cagcgatggg ggcggggggg gggggggggg gggcccgggg gggggggggg
481 gggggggggg gggggggggg gggggggggg gggggggggg gcggcggcgg ccaatcagag
541 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
601 gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc
661 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
721 gcgggacggc ccttctcctc cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg
781 atggttggtt ggtggggtat taatgtttaa ttacctggag cacctgcctg aaatcacttt
841 ttttcaggtt ggaccggtgc caccatggac tataaggacc acgacggaga ctacaaggat
901 catgatattg attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc
961 ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc
1021 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
1081 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
1141 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
1201 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
1261 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
1321 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
1381 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
1441 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
1501 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
1561 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
1621 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
1681 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
1741 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
1801 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
1861 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
1921 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
1981 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
2041 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
2101 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
2161 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
2221 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
2281 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
2341 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
2401 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
2461 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
2521 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
2581 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
2641 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
2701 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
2761 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
2821 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
2881 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
2941 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
3001 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
3061 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
3121 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
3181 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
3241 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
3301 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
3361 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
3421 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
3481 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
3541 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
3601 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
3661 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
3721 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
3781 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
3841 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
3901 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
3961 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
4021 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
4081 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
4141 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
4201 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
4261 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
4321 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
4381 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
4441 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
4501 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
4561 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
4621 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
4681 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
4741 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
4801 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
4861 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
4921 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
4981 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
5041 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaaaag gccggcggcc
5101 acgaaaaagg ccggccaggc aaaaaagaaa aagtaagaat tcctagagct cgctgatcag
5161 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
5221 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
5281 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
5341 aggattggga agagaatagc aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg
5401 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
5461 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
5521 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
5581 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
5641 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
5701 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
5761 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
5821 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
5881 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
5941 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
6001 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
6061 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
6121 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
6181 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
6241 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
6301 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
6361 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
6421 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
6481 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
6541 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
6601 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
6661 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
6721 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
6781 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
6841 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
6901 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
6961 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
7021 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
7081 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
7141 agccggtgag cgtggaagcc gcggtatcat tgcagcactg gggccagatg gtaagccctc
7201 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
7261 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
7321 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
7381 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
7441 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
7501 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
7561 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
7621 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
7681 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
7741 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
7801 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
7861 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
7921 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
7981 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
8041 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
8101 ctggcctttt gctcacatgt gagggcctat ttcccatgat tccttcatat ttgcatatac
8161 gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag
8221 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
8281 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
8341 tatatatctt gtggaaagga cgaaacaccg ggtcttcgag aagacctgtt ttagagctag
8401 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
8461 tgcttttttg ttttagagct agaaatagca agttaaaata aggctagtcc gtttttagcg
8521 cgtgcgccaa ttctgcagac aaatggctct agaggcatgt gagggcctat ttcccatgat
8581 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac
8641 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
8701 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
8761 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg catcgtacgc
8821 gtacgtgttg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
8881 tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag ctagaaatag caagttaaaa
8941 taaggctagt c
//
