| 质粒名称: | LentiGuide Cherry |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 170510 |
| 存储实验室: | Pavel Tolar |
|
载体骨架基本信息 |
|
| 载体名称: | lentiGuide-Puro |
| 载体出品公司: | Feng Zhang lab |
| 完整质粒大小: | 10277 bp |
| 载体类型: | 哺乳细胞表达、Lentiviral, CRISPR |
| 筛选标记: | mCherry |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称1: | S. pyogenes sgRNA cassette / S. pyogenes CRISPR customizable RNA element |
| 物种来源: | Synthetic |
| 插入大小: | 100bp |
| 启动子: | hU6 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BsmBI(未破坏) |
| 3’克隆位点: | BsmBI (未破坏) |
| 5’测序引物： | ACTATCATATGCTTACCGTAAC |
| 插入基因名称2: | mCherry |
| 物种来源: | Synthetic |
| 插入大小: | 698bp |
| 启动子: | EF-1a |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BsiWI (not destroyed) |
| 3’克隆位点: | MluI (not destroyed) |
| 5’测序引物： | tggttcattctcaagcctcag |
|
文献引用方法 |
|
| 材料和方法部分: |
LentiGuide Cherry was a gift from Pavel Tolar (Addgene plasmid # 170510 ; http://n2t.net/addgene:170510 ; RRID:Addgene_170510)
|
| 参考文献部分: | Genome-wide screens identify calcium signaling as a key regulator of IgE+ plasma cell differentiation and survival. Newman R, Tolar P. biorXiv 2021.03.02.433398 10.1101/2021.03.02.433398 |
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 LentiGuide Cherry 10277 bp ds-DNA circular SYN 27-SEP-2021
DEFINITION Expresses S. pyogenes CRISPR chimeric RNA element with customizable
sgRNA from U6 promoter and mCherry marker from EF-1a promoter. 3rd
generation lentiviral backbone..
ACCESSION .
VERSION .
KEYWORDS LentiGuide Cherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10277)
AUTHORS Newman R, Tolar P
TITLE Genome-wide screens identify calcium signaling as a key regulator of
IgE+ plasma cell differentiation and survival
JOURNAL biorXiv 2021.03.02.433398
REFERENCE 2 (bases 1 to 10277)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 27, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10277
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 87..1265
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 318..1256
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1213..1233
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
primer_bind complement(1406..1424)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1674..1693
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 1987..2575
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2040..2060)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2458..2469)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 2647..2880
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 2958..3079
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2995..3014)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3049..3068
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 3119..3254
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3181..3200
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(3274..3293)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3275..3293)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3303..3320)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3303..3319)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3312..3334)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3461..3916
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3548..3567)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3758..3779
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3942..4046
/gene="bla"
/label=AmpR promoter
CDS 4047..4907
/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(4265..4284)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5078..5666
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5567..5586
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5820..5837
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5954..5975
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5990..6020
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6028..6044
/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)."
primer_bind 6033..6055
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6052..6068
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6052..6068
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 6087..6107
/label=T3
/note="T3 promoter, forward primer"
promoter 6089..6107
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 6135..6361
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 6362..6542
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 6589..6714
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 7207..7440
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7625..7669
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
promoter 7847..8087
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 7847..7867
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 8018..8037
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 9977..10052
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
misc_feature 10108..10225
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
ORIGIN
1 cccgggtgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
361 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
421 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
481 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
541 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
601 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
661 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
721 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
781 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
841 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
901 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
961 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
1021 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
1081 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
1141 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
1201 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
1261 cgtgacgtac gcctcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
1321 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
1381 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
1441 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
1501 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
1561 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
1621 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
1681 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
1741 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
1801 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
1861 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
1921 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta aacgcgttaa
1981 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
2041 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
2101 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
2161 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
2221 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
2281 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
2341 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
2401 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
2461 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
2521 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga
2581 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
2641 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
2701 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
2761 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
2821 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
2881 gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
2941 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
3001 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
3061 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat
3121 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
3181 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
3241 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc
3301 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
3361 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
3421 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
3481 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
3541 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
3601 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
3661 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
3721 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
3781 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
3841 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
3901 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
3961 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
4021 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
4081 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
4141 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
4201 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
4261 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
4321 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
4381 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
4441 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
4501 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
4561 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
4621 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
4681 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
4741 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
4801 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
4861 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
4921 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
4981 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
5041 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
5101 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
5161 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
5221 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
5281 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
5341 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
5401 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
5461 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
5521 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
5581 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
5641 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
5701 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
5761 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
5821 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
5881 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
5941 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
6001 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
6061 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
6121 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
6181 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
6241 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
6301 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa
6361 cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
6421 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
6481 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
6541 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
6601 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
6661 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
6721 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
6781 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
6841 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
6901 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
6961 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
7021 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
7081 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
7141 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
7201 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
7261 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
7321 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
7381 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
7441 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
7501 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
7561 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
7621 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
7681 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
7741 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
7801 accattatcg tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc
7861 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa
7921 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
7981 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
8041 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggaga
8101 cggttgtaaa tgagcacaca aaatacacat gctaaaatat tatattctat gacctttata
8161 aaatcaacca aaatcttctt tttaataact ttagtatcaa taattagaat ttttatgttc
8221 ctttttgcaa acttttaata aaaatgagca aaataaaaaa acgctagttt tagtaactcg
8281 cgttgttttc ttcaccttta ataatagcta ctccaccact tgttcctaag cggtcagctc
8341 ctgcttcaat cattttttga gcatcttcaa atgttctaac tccaccagct gctttaacta
8401 aagcattgtc tttaacaact gacttcatta gtttaacatc ttcaaatgtt gcacctgatt
8461 ttgaaaatcc tgttgatgtt ttaacaaatt ctaatccagc ttcaacagct atttcacaag
8521 ctttcatgat ttcttctttt gttaataaac aattttccat aatacattta acaacatgtg
8581 atccagctgc tttttttaca gctttcatgt cttctaaaac taattcataa tttttgtctt
8641 ttaatgcacc aatatttaat accatatcaa tttctgttgc accatcttta attgcttcag
8701 aaacttcgaa tgcttttgta gctgttgtgc atgcacctag aggaaaacct acaacatttg
8761 ttattcctac atttgtgcct tttaataatt ctttacaata gcttgttcaa tatgaattaa
8821 cacaaactgt tgcaaaatca aattcaattg cttcatcaca taattgttta atttcagctt
8881 tcgtagcatc ttgttttaat aatgtgtgat ctatatattt gtttagtttc attttttctc
8941 ctatatattc atttttaatt ttaattcttt aataatttcg tctactttaa ctttagcgtt
9001 ttgaacagat tcaccaacac ctataaaata aatttttagt ttaggttcag ttccacttgg
9061 gcgaacagca aatcatgact tatcttctaa ataaaatttt agtaagtctt gtcctggcat
9121 attatacatt ccatcgatgt agtcttcaac attaacaact ttaagtccag caatttgagt
9181 taagggtgtt gctctcaatg atttcattaa tggttcaatt tttaatttct tttcttctgg
9241 tttaaaattc aagtttaaag tgaaagtgta atatgcaccc atttctttaa ataaatcttc
9301 taaatagtct actaatgttt tattttgttt tttataaaat caagcagcct ctgctattaa
9361 tatagaagct tgtattccat ctttatctct agctgagtca tcaattacat atccataact
9421 ttcttcataa gcaaaaacaa aatttaatcc gttatcttct tctttagcaa tttctctacc
9481 cattcattta aatccagtta aagtttttac aatattaact ccatattttt catgagcgat
9541 tctatcaccc aaatcacttg ttacaaaact tgaatataga gccggatttt ttggaatgct
9601 atttaagcgt tttagatttg ataattttca atcaattaaa attggtcctg tttgatttcc
9661 atctaatctt acaaaatgac catcatgttt tattgccatt ccaaatctgt cagcatctgg
9721 gtcattcata ataataatat ctgcatcatg tttaatacca tattcaagcg gtatttttca
9781 tgcaggatca aattctggat ttggatttac aacattttta aatgtttcat cttcaaatgc
9841 atgctcttca acctcaataa cgttatatcc tgattcacgt aatatttttg gggtaaattt
9901 agttcctgtt ccattaactg cgctaaaaat aatttttaaa tcttttttag cttcttgctc
9961 ttttttgtac gtctctgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
10021 atcaacttga aaaagtggca ccgagtcggt gcttttttaa gcttggcgta actagatctt
10081 gagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
10141 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
10201 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccactttg
10261 gcgccggctc gaggggg
//
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