| 质粒名称: | LentiGuide_DKO |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 183193 |
| 存储实验室: | Lang Li |
|
载体骨架基本信息 |
|
| 载体名称: | lentiGuide-Puro |
| 载体出品公司: | Addgene Plasmid #52963 |
| 空载体大小: | 10183bp |
| 完整质粒大小: | 8789bp |
| 载体类型: | 哺乳细胞表达, Lentiviral, CRISPR |
| 筛选标记: | Puromycin |
| 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 |
|
| 启动子: | U6, mU6 |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | hU6-F |
|
文献引用方法 |
|
| 材料和方法部分: | LentiGuide_DKO was a gift from Lang Li (Addgene plasmid # 183193 ; http://n2t.net/addgene:183193 ; RRID:Addgene_183193) |
| 参考文献部分: | Generation of dual-gRNA library for combinatorial CRISPR screening of synthetic lethal gene pairs. Tang S, Wu X, Liu J, Zhang Q, Wang X, Shao S, Gokbag B, Fan K, Liu X, Li F, Cheng L, Li L. STAR Protoc. 2022 Aug 24;3(3):101556. doi: 10.1016/j.xpro.2022.101556. eCollection 2022 Sep 16. 10.1016/j.xpro.2022.101556 PubMed 36060092 |
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_DKO 8789 bp ds-DNA circular SYN 21-APR-2022
DEFINITION Plasmid with two U6 promoters and two gRNA scaffolds that allows for
inserting two gRNAs for combinatorial CRISPR Screen or
double-knock-out (DKO) screen.
ACCESSION .
VERSION .
KEYWORDS LentiGuide_DKO
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8789)
TITLE Fast and Efficient Generation of Paired-gRNA Library Facilitates
Combinatorial CRISPR Screen for Discovering Synthetic Lethal Gene
Pairs
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8789)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 21, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8789
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 162..402
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 162..182
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 333..352
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
promoter 546..859
/label=U6 promoter
/note="RNA polymerase III promoter for mouse U6 snRNA (Das
et al., 1988)"
primer_bind 778..798
/label=mU6-F
/note="Mouse U6 promoter, forward primer"
misc_RNA 898..973
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
misc_feature 1029..1146
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1285..2463
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 1516..2454
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 2411..2431
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 2476..3075
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(2476..2495)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2972..2992
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 3091..3679
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3144..3164)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3562..3573)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 3751..3984
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4062..4183
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4099..4118)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4153..4172
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4223..4358
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4285..4304
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4378..4397)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4379..4397)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4407..4424)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4407..4423)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4416..4438)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4565..5020
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4652..4671)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4862..4883
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5046..5150
/gene="bla"
/label=AmpR promoter
CDS 5151..6011
/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(5369..5388)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6182..6770
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6671..6690
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6924..6941
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7058..7079
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7094..7124
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7132..7148
/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 7137..7159
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7156..7172
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7156..7172
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 7191..7211
/label=T3
/note="T3 promoter, forward primer"
promoter 7193..7211
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 7239..7465
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 7466..7646
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 7693..7818
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8311..8544
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 8729..8773
/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"
ORIGIN
1 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
61 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
121 tatcgtttca gacccacctc ccaaccccga ggggacccag agagggccta tttcccatga
181 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga
241 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
301 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
361 agtatttcga tttcttggct ttatatatct tgtggaaagg actctagaac caggatgggc
421 accacccgtt taagagctaa gctggaaaca gcatagcaag tttaaataag gctagtccgt
481 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt gtactgagtc gcccagtctc
541 agatagatcc gacgccgcca tctctaggcc cgcgccggcc ccctcgcaca gacttgtggg
601 agaagctcgg ctactcccct gccccggtta atttgcatat aatatttcct agtaactata
661 gaggcttaat gtgcgataaa agacagataa tctgttcttt ttaatactag ctacatttta
721 catgataggc ttggatttct ataagagata caaatactaa attattattt taaaaaacag
781 cacaaaagga aactcaccct aactgtaaag taattgtgtg ttttgagact ataaatatcc
841 cttggagaaa agccttgttc accggagacg gcttcttgct cttttttgta cgtctctgtt
901 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
961 accgagtcgg tgctttttta agcttggcgt aactagatct tgagacaaat ggcagtattc
1021 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
1081 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
1141 aattttcggg tttattacag ggacagcaga gatccacttt ggcgccggct cgagggggcc
1201 cgggtgcaaa gatggataaa gttttaaaca gagaggaatc tttgcagcta atggaccttc
1261 taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg
1321 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc ctagagaagg
1381 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
1441 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
1501 gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt
1561 tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc
1621 gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg agccccttcg
1681 cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg
1741 caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga
1801 cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatctgcac
1861 actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca
1921 tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa
1981 gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg
2041 gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct
2101 gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc
2161 acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac
2221 cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc gtctttaggt
2281 tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt
2341 aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct
2401 tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg
2461 tgacgtacgg ccaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
2521 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
2581 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
2641 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc cgtggcggtc
2701 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
2761 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
2821 caccggccca aggagcccgc gtggttcctg gccaccgtcg gagtctcgcc cgaccaccag
2881 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
2941 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
3001 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
3061 aagcccggtg cctgaacgcg ttaagtcgac aatcaacctc tggattacaa aatttgtgaa
3121 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
3181 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
3241 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
3301 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
3361 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
3421 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
3481 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
3541 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
3601 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
3661 ctttgggccg cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc
3721 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
3781 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
3841 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
3901 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
3961 agtcagtgtg gaaaatctct agcagtacgt atagtagttc atgtcatctt attattcagt
4021 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
4081 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
4141 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
4201 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
4261 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
4321 gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc
4381 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
4441 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
4501 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
4561 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
4621 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
4681 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
4741 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
4801 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
4861 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
4921 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
4981 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
5041 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
5101 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
5161 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
5221 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
5281 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
5341 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
5401 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
5461 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
5521 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
5581 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
5641 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
5701 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
5761 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
5821 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
5881 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
5941 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
6001 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
6061 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
6121 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
6181 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
6241 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
6301 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
6361 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
6421 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
6481 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
6541 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
6601 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
6661 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
6721 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
6781 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
6841 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
6901 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
6961 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
7021 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
7081 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
7141 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
7201 ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt
7261 gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca
7321 ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca
7381 gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt
7441 taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga
7501 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
7561 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
7621 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
7681 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
7741 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
7801 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
7861 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
7921 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
7981 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
8041 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
8101 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
8161 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
8221 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
8281 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
8341 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
8401 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
8461 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
8521 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
8581 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
8641 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
8701 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
8761 atgggcaagt ttgtggaatt ggtttaaca
//
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