| 质粒名称: | pDY1052 PASTEv3 pCMV-SpCas9-XTEN3-RT(L139P)-(GGS)6-BxbINT |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 179105 |
| 存储实验室: | Omar Abudayyeh, Jonathan Gootenberg |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 11428 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，CRISPR |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | pCMV-SpCas9-XTEN3-RT(L139P)-(GGS)6-BxbINT |
|
文献引用方法 |
|
| 材料和方法部分: | pDY1052 PASTEv3 pCMV-SpCas9-XTEN3-RT(L139P)-(GGS)6-BxbINT was a gift from Omar Abudayyeh & Jonathan Gootenberg (Addgene plasmid # 179105 ; http://n2t.net/addgene:179105 ; RRID:Addgene_179105) |
| 参考文献部分: | Drag-and-drop genome insertion of large sequences without double-strand DNA cleavage using CRISPR-directed integrases. Yarnall MTN, Ioannidi EI, Schmitt-Ulms C, Krajeski RN, Lim J, Villiger L, Zhou W, Jiang K, Garushyants SK, Roberts N, Zhang L, Vakulskas CA, Walker JA 2nd, Kadina AP, Zepeda AE, Holden K, Ma H, Xie J, Gao G, Foquet L, Bial G, Donnelly SK, Miyata Y, Radiloff DR, Henderson JM, Ujita A, Abudayyeh OO, Gootenberg JS. Nat Biotechnol. 2022 Nov 24. doi: 10.1038/s41587-022-01527-4. 10.1038/s41587-022-01527-4 PubMed 36424489 |
pDY1052 PASTEv3 pCMV-SpCas9-XTEN3-RT(L139P)-(GGS)6-BxbINT
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
-(GGS)6-BxbINT质粒图谱.png)
载体简介
载体序列
LOCUS pDY1052 PASTEv3 pCMV-SpCas9-XTEN3-RT(L139P)-(GGS)6-BxbINT 11428 bp ds-DNA circular SYN 10-FEB-2022
DEFINITION pCMV-SpCas9-XTEN3-RT(L139P)-(GGS)6-BxbINT.
ACCESSION .
VERSION .
KEYWORDS pDY1052 PASTEv3 pCMV-SpCas9-XTEN3-RT(L139P)-(GGS)6-BxbINT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11428)
AUTHORS Ioannidi EI, Yarnall MTN, Schmitt-Ulms C, Krajeski RN, Lim J,
Villiger L, Zhou W, Jiang K, Roberts N, Zhang L, Vakulskas CA,
Walker II JA, Kadina AP, Zepeda AE, Holden K, Gootenberg JS,
Abudayyeh OO
TITLE Drag-and-drop genome insertion without DNA cleavage with
CRISPR-directed integrases
JOURNAL bioRxiv 2021.11.01.466786
REFERENCE 2 (bases 1 to 11428)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 10, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..11428
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(70..89)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 261..640
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 641..844
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 795..815
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 886..905
/label=T7
/note="T7 promoter, forward primer"
promoter 886..904
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 954..974
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 975..5075
/codon_start=1
/product="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
/label=Cas9(H840A)
/note="generates RNA-guided single strand nicks in DNA due
to the H840A mutation in the HNH catalytic domain"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 7344..7364
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7419..7439
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7440..7466
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
primer_bind complement(8992..9009)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 8998..9222
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(9293..9309)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9293..9309)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(9306..9328)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 9317..9333
/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(9341..9371)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9386..9407
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(9524..9541)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9695..10283)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(9775..9794)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(10454..11314)
/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 11077..11096
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(11315..11419)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ccgaaaagtg ccacctgacg tcgacggatc gggagatcga tctcccgatc ccctagggtc
61 tactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
121 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
181 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
241 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
301 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
361 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
421 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
481 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
541 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
601 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
661 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
721 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
781 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
841 agctggttta gtgaaccgtc agatccgcta gagatccgcg gccgctaata cgactcacta
901 tagggagagc cgccaccatg aaacggacag ccgacggaag cgagttcgag tcaccaaaga
961 agaagcggaa agtcgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg
1021 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
1081 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
1141 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
1201 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
1261 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
1321 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
1381 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
1441 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
1501 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
1561 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
1621 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
1681 tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
1741 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
1801 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
1861 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
1921 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
1981 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
2041 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
2101 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
2161 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
2221 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
2281 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
2341 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
2401 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
2461 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
2521 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
2581 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
2641 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
2701 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
2761 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
2821 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
2881 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
2941 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
3001 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
3061 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
3121 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
3181 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
3241 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
3301 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
3361 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
3421 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
3481 atgtggacgc tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
3541 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
3601 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
3661 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
3721 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
3781 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
3841 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
3901 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
3961 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
4021 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
4081 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
4141 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
4201 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
4261 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
4321 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc
4381 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
4441 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
4501 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
4561 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
4621 ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
4681 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
4741 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
4801 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
4861 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
4921 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca
4981 ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
5041 agacacggat cgacctgtct cagctgggag gtgactctgg aggatctagc ggaggatcct
5101 ctggcagcga gacaccagga acaagcgagt cagcaacacc agagagctct ggtagcgaga
5161 cacccggtac cagtgaaagc gccacgccag aaagcagtgg gagtgagact ccgggtacat
5221 ctgaatcagc gacaccggaa tcaagtggcg gcagcagtgg cggcagcagc accctaaata
5281 tagaagatga gtatcggcta catgagacct caaaagagcc agatgtttct ctagggtcca
5341 catggctgtc tgattttcct caggcctggg cggaaaccgg gggcatggga ctggcagttc
5401 gccaagctcc tctgatcata cctctgaaag caacctctac ccccgtgtcc ataaaacaat
5461 accccatgtc acaagaagcc agactgggga tcaagcccca catacagaga ctgttggacc
5521 agggaatact ggtaccctgc cagtccccct ggaacacgcc cctgctaccc gttaagaaac
5581 cagggactaa tgattatagg cctgtccagg atctgagaga agtcaacaag cgggtggaag
5641 acatccaccc caccgtgccc aacccttaca acctcttgag cgggccccca ccgtcccacc
5701 agtggtacac tgtgcttgat ttaaaggatg cctttttctg cctgagactc caccccacca
5761 gtcagcctct cttcgccttt gagtggagag atccagagat gggaatctca ggacaattga
5821 cctggaccag actcccacag ggtttcaaaa acagtcccac cctgtttaat gaggcactgc
5881 acagagacct agcagacttc cggatccagc acccagactt gatcctgcta cagtacgtgg
5941 atgacttact gctggccgcc acttctgagc tagactgcca acaaggtact cgggccctgt
6001 tacaaaccct agggaacctc gggtatcggg cctcggccaa gaaagcccaa atttgccaga
6061 aacaggtcaa gtatctgggg tatcttctaa aagagggtca gagatggctg actgaggcca
6121 gaaaagagac tgtgatgggg cagcctactc cgaagacccc tcgacaacta agggagttcc
6181 tagggaaggc aggcttctgt cgcctcttca tccctgggtt tgcagaaatg gcagcccccc
6241 tgtaccctct caccaaaccg gggactctgt ttaattgggg cccagaccaa caaaaggcct
6301 atcaagaaat caagcaagct cttctaactg ccccagccct ggggttgcca gatttgacta
6361 agccctttga actctttgtc gacgagaagc agggctacgc caaaggtgtc ctaacgcaaa
6421 aactgggacc ttggcgtcgg ccggtggcct acctgtccaa aaagctagac ccagtagcag
6481 ctgggtggcc cccttgccta cggatggtag cagccattgc cgtactgaca aaggatgcag
6541 gcaagctaac catgggacag ccactagtca ttctggcccc ccatgcagta gaggcactag
6601 tcaaacaacc ccccgaccgc tggctttcca acgcccggat gactcactat caggccttgc
6661 ttttggacac ggaccgggtc cagttcggac cggtggtagc cctgaacccg gctacgctgc
6721 tcccactgcc tgaggaaggg ctgcaacaca actgccttga tatcctggcc gaagcccacg
6781 gaacccgacc cgacctaacg gaccagccgc tcccagacgc cgaccacacc tggtacacgg
6841 atggaagcag tctcttacaa gagggacagc gtaaggcggg agctgcggtg accaccgaga
6901 ccgaggtaat ctgggctaaa gccctgccag ccgggacatc cgctcagcgg gctgaactga
6961 tagcactcac ccaggcccta aagatggcag aaggtaagaa gctaaatgtt tatactgata
7021 gccgttatgc ttttgctact gcccatatcc atggagaaat atacagaagg cgtgggtggc
7081 tcacatcaga aggcaaagag atcaaaaata aagacgagat cttggcccta ctaaaagccc
7141 tctttctgcc caaaagactt agcataatcc attgtccagg acatcaaaag ggacacagcg
7201 ccgaggctag aggcaaccgg atggctgacc aagcggcccg aaaggcagcc atcacagaga
7261 ctccagacac ctctaccctc ctcatagaaa attcatcacc ctctggcggc tcaaaaagaa
7321 ccgccgacgg cagcgaattc gagcccaaga agaagaggaa agtcgggggg tcaggtggat
7381 ccggcggaag tggcggatcc ggtggatctg gcggcagtcc aaaaaagaaa agaaaagtgt
7441 atccctatga tgtccccgat tatgccggtt caagagccct ggtcgtgatt agactgagcc
7501 gagtgacaga cgccaccaca agtcccgaga gacagctgga atcatgccag cagctctgtg
7561 ctcagcgggg ttgggatgtg gtcggcgtgg cagaggatct ggacgtgagc ggggccgtcg
7621 atccattcga cagaaagagg aggcccaacc tggcaagatg gctcgctttc gaggaacagc
7681 cctttgatgt gatcgtcgcc tacagagtgg accggctgac ccgctcaatt cgacatctcc
7741 agcagctggt gcattgggct gaggaccaca agaaactggt ggtcagcgca acagaagccc
7801 acttcgatac taccacacct tttgccgctg tggtcatcgc actgatgggc actgtggccc
7861 agatggagct cgaagctatc aaggagcgaa acaggagcgc agcccatttc aatattaggg
7921 ccggtaaata cagaggctcc ctgccccctt ggggatatct ccctaccagg gtggatgggg
7981 agtggagact ggtgccagac cccgtccaga gagagcggat tctggaagtg taccacagag
8041 tggtcgataa ccacgaacca ctccatctgg tggcacacga cctgaataga cgcggcgtgc
8101 tctctccaaa ggattatttt gctcagctgc agggaagaga gccacaggga agagaatgga
8161 gtgctactgc actgaagaga tctatgatca gtgaggctat gctgggttac gcaacactca
8221 atggcaaaac tgtccgggac gatgacggag cccctctggt gagggctgag cctattctca
8281 ccagagagca gctcgaagct ctgcgggcag aactggtcaa gactagtcgc gccaaacctg
8341 ccgtgagcac cccaagcctg ctcctgaggg tgctgttctg cgccgtctgt ggagagccag
8401 catacaagtt tgccggcgga gggcgcaaac atccccgcta tcgatgcagg agcatggggt
8461 tccctaagca ctgtggaaac gggacagtgg ccatggctga gtgggacgcc ttttgcgagg
8521 aacaggtgct ggatctcctg ggtgacgctg agcggctgga aaaagtgtgg gtggcaggat
8581 ctgactccgc tgtggagctg gcagaagtca atgccgagct cgtggatctg acttccctca
8641 tcggatctcc tgcatataga gctgggtccc cacagagaga agctctggac gcacgaattg
8701 ctgcactcgc tgctagacag gaggaactgg agggcctgga ggccaggccc tctggatggg
8761 agtggcgaga aaccggacag aggtttgggg attggtggag ggagcaggac accgcagcca
8821 agaacacatg gctgagatcc atgaatgtcc ggctcacatt cgacgtgcgc ggtggcctga
8881 ctcgaaccat cgattttggc gacctgcagg agtatgaaca gcacctgaga ctggggtccg
8941 tggtcgaaag actgcacact gggatgtcct aggtttaaac ccgctgatca gcctcgactg
9001 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
9061 aaggtgccac tcccactgtc ctttcctaat aaaatgagaa aattgcatcg cattgtctga
9121 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
9181 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
9241 ccagctgggg ctcgataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
9301 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
9361 taaagtgtaa agcctagggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
9421 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
9481 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
9541 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
9601 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
9661 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
9721 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
9781 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
9841 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
9901 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
9961 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
10021 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
10081 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
10141 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
10201 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
10261 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacactc
10321 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
10381 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
10441 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
10501 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
10561 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
10621 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
10681 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
10741 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
10801 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
10861 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
10921 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
10981 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
11041 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
11101 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
11161 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
11221 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
11281 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
11341 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
11401 aacaaatagg ggttccgcgc acatttcc
//
