| 质粒名称: | pC0046-EF1a-PspCas13b-NES-HIV |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 103862 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | EF1a-lenti |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 9000 bp |
| 完整质粒大小: | 12606 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒，CRISPR |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | PspCas13b |
| 物种来源: | Prevotella sp. P5-125 |
| 插入大小: | 3270 bp |
| 基因突变: | -- |
| GenBank编号 | WP_044065294 |
| 启动子 | -- |
| 标签/融合蛋白: |
HIV NES（插入件上的C端子）
3xHA（插入件上的C端子）
|
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pC0046-EF1a-PspCas13b-NES-HIV was a gift from Feng Zhang (Addgene plasmid # 103862 ; http://n2t.net/addgene:103862 ; RRID:Addgene_103862) |
| 参考文献部分: | RNA editing with CRISPR-Cas13. Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung J, Zhang F. Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180. 10.1126/science.aaq0180 PubMed 29070703 |
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
载体简介
载体序列
LOCUS pC0046-EF1a-PspCas13b-NES-HIV 12607 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses PspCas13b in mammalian cells for knockdown of target RNAs
in combination with compatible guide RNAs. Localized to the
cytoplasm.
ACCESSION .
VERSION .
KEYWORDS pC0046-EF1a-PspCas13b-NES-HIV
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12607)
AUTHORS Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung
J, Zhang F
TITLE RNA editing with CRISPR-Cas13.
JOURNAL Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180.
PUBMED 29070703
REFERENCE 2 (bases 1 to 12607)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..12607
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 382..580
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 598..778
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 825..950
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1443..1676
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2203..2320
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2445..3623
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2676..3614
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 3571..3591
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 6918..6944
/codon_start=1
/product="nuclear export signal from the HIV Rev protein
(Fischer et al., 1995)"
/label=NES
/translation="LPPLERLTL"
CDS 6954..7034
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
misc_feature 7062..7650
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(7115..7135)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(7533..7544)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(7653..7669)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7654..7670)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 8175..8355
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(8381..8398)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 8387..8611
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 8657..9085
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8744..8763)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8954..8975
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(9094..9114)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 9099..9428
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 9279..9414
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 9341..9360
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 9476..9523
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 9542..9916
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 10046..10167
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(10083..10102)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 10137..10156
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(10216..10232)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10216..10232)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(10229..10251)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 10240..10256
/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(10264..10294)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 10309..10330
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(10447..10464)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(10618..11206)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(10698..10717)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(11377..12237)
/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 12000..12019
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(12238..12342)
/gene="bla"
/label=AmpR promoter
primer_bind complement(12417..12436)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
2341 cagagatcca gtttggttaa ttaatgcaaa gatggataaa gttttaaaca gagaggaatc
2401 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
2461 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
2521 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
2581 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
2641 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg
2701 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc
2761 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt
2821 gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc
2881 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag
2941 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa
3001 atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg
3061 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa
3121 tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt
3181 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa
3241 gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag
3301 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt
3361 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt
3421 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg
3481 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg
3541 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt
3601 ttcttccatt tcaggtgtcg tgacgtacgg ccaccatgaa catccccgct ctggtggaaa
3661 accagaagaa gtactttggc acctacagcg tgatggccat gctgaacgct cagaccgtgc
3721 tggaccacat ccagaaggtg gccgatattg agggcgagca gaacgagaac aacgagaatc
3781 tgtggtttca ccccgtgatg agccacctgt acaacgccaa gaacggctac gacaagcagc
3841 ccgagaaaac catgttcatc atcgagcggc tgcagagcta cttcccattc ctgaagatca
3901 tggccgagaa ccagagagag tacagcaacg gcaagtacaa gcagaaccgc gtggaagtga
3961 acagcaacga catcttcgag gtgctgaagc gcgccttcgg cgtgctgaag atgtacaggg
4021 acctgaccaa ccactacaag acctacgagg aaaagctgaa cgacggctgc gagttcctga
4081 ccagcacaga gcaacctctg agcggcatga tcaacaacta ctacacagtg gccctgcgga
4141 acatgaacga gagatacggc tacaagacag aggacctggc cttcatccag gacaagcggt
4201 tcaagttcgt gaaggacgcc tacggcaaga aaaagtccca agtgaatacc ggattcttcc
4261 tgagcctgca ggactacaac ggcgacacac agaagaagct gcacctgagc ggagtgggaa
4321 tcgccctgct gatctgcctg ttcctggaca agcagtacat caacatcttt ctgagcaggc
4381 tgcccatctt ctccagctac aatgcccaga gcgaggaacg gcggatcatc atcagatcct
4441 tcggcatcaa cagcatcaag ctgcccaagg accggatcca cagcgagaag tccaacaaga
4501 gcgtggccat ggatatgctc aacgaagtga agcggtgccc cgacgagctg ttcacaacac
4561 tgtctgccga gaagcagtcc cggttcagaa tcatcagcga cgaccacaat gaagtgctga
4621 tgaagcggag cagcgacaga ttcgtgcctc tgctgctgca gtatatcgat tacggcaagc
4681 tgttcgacca catcaggttc cacgtgaaca tgggcaagct gagatacctg ctgaaggccg
4741 acaagacctg catcgacggc cagaccagag tcagagtgat cgagcagccc ctgaacggct
4801 tcggcagact ggaagaggcc gagacaatgc ggaagcaaga gaacggcacc ttcggcaaca
4861 gcggcatccg gatcagagac ttcgagaaca tgaagcggga cgacgccaat cctgccaact
4921 atccctacat cgtggacacc tacacacact acatcctgga aaacaacaag gtcgagatgt
4981 ttatcaacga caaagaggac agcgccccac tgctgcccgt gatcgaggat gatagatacg
5041 tggtcaagac aatccccagc tgccggatga gcaccctgga aattccagcc atggccttcc
5101 acatgtttct gttcggcagc aagaaaaccg agaagctgat cgtggacgtg cacaaccggt
5161 acaagagact gttccaggcc atgcagaaag aagaagtgac cgccgagaat atcgccagct
5221 tcggaatcgc cgagagcgac ctgcctcaga agatcctgga tctgatcagc ggcaatgccc
5281 acggcaagga tgtggacgcc ttcatcagac tgaccgtgga cgacatgctg accgacaccg
5341 agcggagaat caagagattc aaggacgacc ggaagtccat tcggagcgcc gacaacaaga
5401 tgggaaagag aggcttcaag cagatctcca caggcaagct ggccgacttc ctggccaagg
5461 acatcgtgct gtttcagccc agcgtgaacg atggcgagaa caagatcacc ggcctgaact
5521 accggatcat gcagagcgcc attgccgtgt acgatagcgg cgacgattac gaggccaagc
5581 agcagttcaa gctgatgttc gagaaggccc ggctgatcgg caagggcaca acagagcctc
5641 atccatttct gtacaaggtg ttcgcccgca gcatccccgc caatgccgtc gagttctacg
5701 agcgctacct gatcgagcgg aagttctacc tgaccggcct gtccaacgag atcaagaaag
5761 gcaacagagt ggatgtgccc ttcatccggc gggaccagaa caagtggaaa acacccgcca
5821 tgaagaccct gggcagaatc tacagcgagg atctgcccgt ggaactgccc agacagatgt
5881 tcgacaatga gatcaagtcc cacctgaagt ccctgccaca gatggaaggc atcgacttca
5941 acaatgccaa cgtgacctat ctgatcgccg agtacatgaa gagagtgctg gacgacgact
6001 tccagacctt ctaccagtgg aaccgcaact accggtacat ggacatgctt aagggcgagt
6061 acgacagaaa gggctccctg cagcactgct tcaccagcgt ggaagagaga gaaggcctct
6121 ggaaagagcg ggcctccaga acagagcggt acagaaagca ggccagcaac aagatccgca
6181 gcaaccggca gatgagaaac gccagcagcg aagagatcga gacaatcctg gataagcggc
6241 tgagcaacag ccggaacgag taccagaaaa gcgagaaagt gatccggcgc tacagagtgc
6301 aggatgccct gctgtttctg ctggccaaaa agaccctgac cgaactggcc gatttcgacg
6361 gcgagaggtt caaactgaaa gaaatcatgc ccgacgccga gaagggaatc ctgagcgaga
6421 tcatgcccat gagcttcacc ttcgagaaag gcggcaagaa gtacaccatc accagcgagg
6481 gcatgaagct gaagaactac ggcgacttct ttgtgctggc tagcgacaag aggatcggca
6541 acctgctgga actcgtgggc agcgacatcg tgtccaaaga ggatatcatg gaagagttca
6601 acaaatacga ccagtgcagg cccgagatca gctccatcgt gttcaacctg gaaaagtggg
6661 ccttcgacac ataccccgag ctgtctgcca gagtggaccg ggaagagaag gtggacttca
6721 agagcatcct gaaaatcctg ctgaacaaca agaacatcaa caaagagcag agcgacatcc
6781 tgcggaagat ccggaacgcc ttcgatcaca acaattaccc cgacaaaggc gtggtggaaa
6841 tcaaggccct gcctgagatc gccatgagca tcaagaaggc ctttggggag tacgccatca
6901 tgaagggatc ccttcaactg cctccacttg aaagactgac actgggtagt tcctacccat
6961 acgatgttcc agattacgct tatccctacg acgtgcctga ttatgcatac ccatatgatg
7021 tccccgacta tgcctaagaa ttcgatatca agcttatcga taatcaacct ctggattaca
7081 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
7141 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
7201 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
7261 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
7321 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
7381 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
7441 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
7501 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
7561 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
7621 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga gacctagaaa
7681 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
7741 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
7801 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
7861 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
7921 gctacttccc tgattggcag aactacacac cagggccagg gatcagatat ccactgacct
7981 ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag
8041 gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
8101 aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc
8161 atccggactg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
8221 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
8281 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
8341 ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
8401 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
8461 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
8521 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
8581 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
8641 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
8701 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
8761 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
8821 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
8881 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
8941 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
9001 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
9061 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
9121 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
9181 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
9241 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
9301 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
9361 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
9421 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
9481 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
9541 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
9601 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
9661 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
9721 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
9781 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
9841 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
9901 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
9961 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
10021 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
10081 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
10141 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
10201 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
10261 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
10321 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
10381 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
10441 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
10501 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
10561 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
10621 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
10681 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
10741 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
10801 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
10861 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
10921 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
10981 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
11041 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
11101 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
11161 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
11221 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
11281 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
11341 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
11401 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
11461 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
11521 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
11581 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
11641 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
11701 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
11761 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
11821 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
11881 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
11941 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
12001 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
12061 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
12121 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
12181 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
12241 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
12301 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
12361 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
12421 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
12481 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
12541 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
12601 acgcgtt
//
