| 质粒名称: | pXR001: EF1a-CasRx-2A-EGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 109049 |
| 存储实验室: | Patrick Hsu |
|
载体骨架基本信息 |
|
| 载体名称: | lenti(AMP) |
| 载体抗性: | 氨苄青霉素 |
| 完整质粒大小: | 12990 bp |
| 载体类型: | 哺乳动物表达，慢病毒，CRISPR |
| 筛选标记: | GFP |
| 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: | CasRx,NLS-RfxCas13d-NLS |
| 物种来源: | H. sapiens (human); Ruminococcus flavefaciens XPD3002 |
| 插入大小: | 3000 bp |
| 启动子 | EF1A |
| 标签/融合蛋白: |
NLS（插入件上的N端子）
NLS（插接件上的C端子）
HA（插入件上的C端子）
2A eGFP（主干上的C端）
|
| 克隆方法: | Gibson克隆 |
| 5' 测序引物 | ggatcttggttcattctcaagcctc |
|
文献引用方法 |
|
| 材料和方法部分: | pXR001: EF1a-CasRx-2A-EGFP was a gift from Patrick Hsu (Addgene plasmid # 109049 ; http://n2t.net/addgene:109049 ; RRID:Addgene_109049) |
| 参考文献部分: | Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors. Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD. Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi: 10.1016/j.cell.2018.02.033. 10.1016/j.cell.2018.02.033 PubMed 29551272 |
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 pXR001: EF1a-CasRx-2A-EGFP 12991 bp ds-DNA circular SYN 06-MAR-2019
DEFINITION CasRx (NLS-RfxCas13d-NLS) with 2A-EGFP for RNA targeting.
ACCESSION .
VERSION .
KEYWORDS pXR001: EF1a-CasRx-2A-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12991)
AUTHORS Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD
TITLE Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR
Effectors.
JOURNAL Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi:
10.1016/j.cell.2018.02.033.
PUBMED 29551272
REFERENCE 2 (bases 1 to 12991)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 6, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..12991
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 61..178
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 303..1481
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 534..1472
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1429..1449
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 1500..1520
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4437..4457
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4467..4493
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 4509..4562
/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 4563..5279
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(4605..4626)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4866..4885)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 5213..5234
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 5304..5892
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5357..5377)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5775..5786)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5895..5911)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5896..5912)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 6417..6597
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(6623..6640)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6629..6853
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 6899..7327
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6986..7005)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7196..7217
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7336..7356)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 7341..7670
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 7521..7656
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 7583..7602
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 7718..7765
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 7784..8158
/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 8288..8409
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8325..8344)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8379..8398
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(8458..8474)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8458..8474)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8471..8493)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8482..8498
/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(8506..8536)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8551..8572
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8689..8706)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8860..9448)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8940..8959)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9619..10479)
/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 10242..10261
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10480..10584)
/gene="bla"
/label=AmpR promoter
primer_bind complement(10659..10678)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 10850..11229
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 11231..11429
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 11384..11404
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 11447..11627
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 11674..11799
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 12292..12525
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
ORIGIN
1 gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa
61 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
121 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
181 ggtttattac agggacagca gagatccagt ttggttaatt aatgcaaaga tggataaagt
241 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
301 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
361 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
421 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
481 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
541 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
601 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
661 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
721 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
781 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
841 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
901 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
961 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
1021 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
1081 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
1141 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
1201 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
1261 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
1321 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
1381 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
1441 cagtggttca aagttttttt cttccatttc aggtgtcgtg acgtacggcc accatgagcc
1501 ccaagaagaa gagaaaggtg gaggccagca tcgaaaaaaa aaagtccttc gccaagggca
1561 tgggcgtgaa gtccacactc gtgtccggct ccaaagtgta catgacaacc ttcgccgaag
1621 gcagcgacgc caggctggaa aagatcgtgg agggcgacag catcaggagc gtgaatgagg
1681 gcgaggcctt cagcgctgaa atggccgata aaaacgccgg ctataagatc ggcaacgcca
1741 aattcagcca tcctaagggc tacgccgtgg tggctaacaa ccctctgtat acaggacccg
1801 tccagcagga tatgctcggc ctgaaggaaa ctctggaaaa gaggtacttc ggcgagagcg
1861 ctgatggcaa tgacaatatt tgtatccagg tgatccataa catcctggac attgaaaaaa
1921 tcctcgccga atacattacc aacgccgcct acgccgtcaa caatatctcc ggcctggata
1981 aggacattat tggattcggc aagttctcca cagtgtatac ctacgacgaa ttcaaagacc
2041 ccgagcacca tagggccgct ttcaacaata acgataagct catcaacgcc atcaaggccc
2101 agtatgacga gttcgacaac ttcctcgata accccagact cggctatttc ggccaggcct
2161 ttttcagcaa ggagggcaga aattacatca tcaattacgg caacgaatgc tatgacattc
2221 tggccctcct gagcggactg aggcactggg tggtccataa caacgaagaa gagtccagga
2281 tctccaggac ctggctctac aacctcgata agaacctcga caacgaatac atctccaccc
2341 tcaactacct ctacgacagg atcaccaatg agctgaccaa ctccttctcc aagaactccg
2401 ccgccaacgt gaactatatt gccgaaactc tgggaatcaa ccctgccgaa ttcgccgaac
2461 aatatttcag attcagcatt atgaaagagc agaaaaacct cggattcaat atcaccaagc
2521 tcagggaagt gatgctggac aggaaggata tgtccgagat caggaaaaat cataaggtgt
2581 tcgactccat caggaccaag gtctacacca tgatggactt tgtgatttat aggtattaca
2641 tcgaagagga tgccaaggtg gctgccgcca ataagtccct ccccgataat gagaagtccc
2701 tgagcgagaa ggatatcttt gtgattaacc tgaggggctc cttcaacgac gaccagaagg
2761 atgccctcta ctacgatgaa gctaatagaa tttggagaaa gctcgaaaat atcatgcaca
2821 acatcaagga atttagggga aacaagacaa gagagtataa gaagaaggac gcccctagac
2881 tgcccagaat cctgcccgct ggccgtgatg tttccgcctt cagcaaactc atgtatgccc
2941 tgaccatgtt cctggatggc aaggagatca acgacctcct gaccaccctg attaataaat
3001 tcgataacat ccagagcttc ctgaaggtga tgcctctcat cggagtcaac gctaagttcg
3061 tggaggaata cgcctttttc aaagactccg ccaagatcgc cgatgagctg aggctgatca
3121 agtccttcgc tagaatggga gaacctattg ccgatgccag gagggccatg tatatcgacg
3181 ccatccgtat tttaggaacc aacctgtcct atgatgagct caaggccctc gccgacacct
3241 tttccctgga cgagaacgga aacaagctca agaaaggcaa gcacggcatg agaaatttca
3301 ttattaataa cgtgatcagc aataaaaggt tccactacct gatcagatac ggtgatcctg
3361 cccacctcca tgagatcgcc aaaaacgagg ccgtggtgaa gttcgtgctc ggcaggatcg
3421 ctgacatcca gaaaaaacag ggccagaacg gcaagaacca gatcgacagg tactacgaaa
3481 cttgtatcgg aaaggataag ggcaagagcg tgagcgaaaa ggtggacgct ctcacaaaga
3541 tcatcaccgg aatgaactac gaccaattcg acaagaaaag gagcgtcatt gaggacaccg
3601 gcagggaaaa cgccgagagg gagaagttta aaaagatcat cagcctgtac ctcaccgtga
3661 tctaccacat cctcaagaat attgtcaata tcaacgccag gtacgtcatc ggattccatt
3721 gcgtcgagcg tgatgctcaa ctgtacaagg agaaaggcta cgacatcaat ctcaagaaac
3781 tggaagagaa gggattcagc tccgtcacca agctctgcgc tggcattgat gaaactgccc
3841 ccgataagag aaaggacgtg gaaaaggaga tggctgaaag agccaaggag agcattgaca
3901 gcctcgagag cgccaacccc aagctgtatg ccaattacat caaatacagc gacgagaaga
3961 aagccgagga gttcaccagg cagattaaca gggagaaggc caaaaccgcc ctgaacgcct
4021 acctgaggaa caccaagtgg aatgtgatca tcagggagga cctcctgaga attgacaaca
4081 agacatgtac cctgttcaga aacaaggccg tccacctgga agtggccagg tatgtccacg
4141 cctatatcaa cgacattgcc gaggtcaatt cctacttcca actgtaccat tacatcatgc
4201 agagaattat catgaatgag aggtacgaga aaagcagcgg aaaggtgtcc gagtacttcg
4261 acgctgtgaa tgacgagaag aagtacaacg ataggctcct gaaactgctg tgtgtgcctt
4321 tcggctactg tatccccagg tttaagaacc tgagcatcga ggccctgttc gataggaacg
4381 aggccgccaa gttcgacaag gagaaaaaga aggtgtccgg caattccgga tccggaccta
4441 agaaaaagag gaaggtggcg gccgcttacc catacgatgt tccagattac gctgctagcg
4501 gcagtggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc
4561 cagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
4621 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
4681 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
4741 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
4801 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
4861 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
4921 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
4981 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
5041 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
5101 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
5161 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
5221 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag
5281 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
5341 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
5401 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
5461 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
5521 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
5581 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
5641 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
5701 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
5761 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
5821 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
5881 ccgcctcccc gcatcgatac cgtcgacctc gagacctaga aaaacatgga gcaatcacaa
5941 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg
6001 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg
6061 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
6121 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc
6181 agaactacac accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc
6241 tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt
6301 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt
6361 ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt
6421 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
6481 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
6541 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg
6601 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
6661 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
6721 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
6781 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
6841 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
6901 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
6961 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
7021 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
7081 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
7141 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
7201 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
7261 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
7321 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
7381 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
7441 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
7501 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
7561 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
7621 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
7681 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
7741 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
7801 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
7861 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
7921 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
7981 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
8041 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
8101 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
8161 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
8221 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
8281 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
8341 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
8401 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
8461 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
8521 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
8581 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
8641 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
8701 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
8761 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
8821 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
8881 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
8941 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
9001 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
9061 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
9121 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
9181 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
9241 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
9301 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
9361 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
9421 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
9481 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
9541 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
9601 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
9661 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
9721 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
9781 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
9841 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
9901 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
9961 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
10021 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
10081 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
10141 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
10201 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
10261 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
10321 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
10381 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
10441 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
10501 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
10561 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
10621 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
10681 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
10741 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
10801 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
10861 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
10921 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
10981 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
11041 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
11101 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
11161 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
11221 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
11281 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
11341 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
11401 cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt
11461 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
11521 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
11581 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
11641 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
11701 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
11761 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
11821 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
11881 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
11941 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
12001 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
12061 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
12121 accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa
12181 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
12241 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
12301 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
12361 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
12421 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
12481 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
12541 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
12601 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
12661 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
12721 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
12781 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
12841 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
12901 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
12961 agagagagac agagacagat ccattcgatt a
//
