| 质粒名称: | pINDUCER21 (ORF-EG) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 46948 |
| 存储实验室: | Stephen Elledge, Thomas Westbrook |
|
载体骨架基本信息 |
|
| 载体名称: | pHAGE-CAGS-W |
| 载体出品公司: | Dr. Richard Mulligan |
| 载体抗性: | 氯霉素和氨苄青霉素 |
| 空载体大小: | 4874 bp |
| 完整质粒大小: | -- |
| 载体修饰: | pINDUCER21（ORF-UG）是用NdeI和SphI消化pINDUCER20制成的。eGFP分别在5′端和3′端用AgeI和SphI进行PCR扩增，然后用AgeI和SphI进行酶切。将载体与eGFP连接，并进行序列验证。pINDUCER20（ORF-UN）是通过用BamHI和SpeI从pSLIK Neo-TGmiR-Luc载体中消化TRE2，并将钝端克隆到SpeI和XhoI消化的噬菌体-CAGS-W载体（Richard Mulligan博士，哈佛医学院，波士顿）的骨干中来产生噬菌体TRE载体。用SpeI从噬菌体EF-DEST载体（G.H.）中切出含有网关受体盒的片段并插入噬菌体TRE中以产生噬菌体TRE-DEST载体。用PacI和ClaI从pSLIK-Neo-TGmiR-Luc载体中切出Ubc-rtTA3-IRES-Neo片段，并将其克隆到SphI和ClaI消化的噬菌体tredest的骨架中。 |
| 载体类型: | 慢病毒载体表达 |
| 筛选标记: | eGFP |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 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: | pINDUCER21 (ORF-EG) was a gift from Stephen Elledge & Thomas Westbrook (Addgene plasmid # 46948 ; http://n2t.net/addgene:46948 ; RRID:Addgene_46948) |
| 参考文献部分: | The pINDUCER lentiviral toolkit for inducible RNA interference in vitro and in vivo. Meerbrey KL, Hu G, Kessler JD, Roarty K, Li MZ, Fang JE, Herschkowitz JI, Burrows AE, Ciccia A, Sun T, Schmitt EM, Bernardi RJ, Fu X, Bland CS, Cooper TA, Schiff R, Rosen JM, Westbrook TF, Elledge SJ. Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3665-70. doi: 10.1073/pnas.1019736108. Epub 2011 Feb 9. 10.1073/pnas.1019736108 PubMed 21307310 |
买家导航
pINDUCER21 (ORF-EG)载体质粒基本信息
pINDUCER21 (ORF-EG)质粒图谱载体图谱和pINDUCER21 (ORF-EG)载体序列质粒序列多克隆位点信息
质粒图谱.png)
pINDUCER21 (ORF-EG)质粒载体简介
pINDUCER21 (ORF-EG)质粒序列载体序列
LOCUS pINDUCER21 (ORF-EG) 12152 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Inducible lentiviral gene expression vector. Chloramphenicol cm +
ccdB cassette may be removed to clone in gene of interest.
ACCESSION .
VERSION .
KEYWORDS pINDUCER21 (ORF-EG)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12152)
AUTHORS Meerbrey KL, Hu G, Kessler JD, Roarty K, Li MZ, Fang JE,
Herschkowitz JI, Burrows AE, Ciccia A, Sun T, Schmitt EM, Bernardi
RJ, Fu X, Bland CS, Cooper TA, Schiff R, Rosen JM, Westbrook TF,
Elledge SJ
TITLE The pINDUCER lentiviral toolkit for inducible RNA interference in
vitro and in vivo.
JOURNAL Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3665-70. doi:
10.1073/pnas.1019736108. Epub 2011 Feb 9.
PUBMED 21307310
REFERENCE 2 (bases 1 to 12152)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..12152
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1721..1765
/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"
misc_feature 2058..2175
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 2230..2500
/label=tetracycline response element
/bound_moiety="tetracycline repressor TetR"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 2533..2571
/label=minimal CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2568..2592
/label=LNCX
/note="Human CMV promoter, forward primer"
protein_bind 2701..2825
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 2850..2880
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 2934..3593
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(3000..3019)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 3935..4240
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 4119..4138
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(4281..4405)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 4458..4484
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter 4704..5882
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 4935..5873
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 5830..5850
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
protein_bind complement(5952..5985)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS 5992..6738
/codon_start=1
/product="improved tetracycline-controlled transactivator"
/label=rtTA-Advanced
/note="In the Tet-On(R) system, rtTA-Advanced binds to the
Tet-responsive element and stimulates transcription only in
the presence of tetracycline or doxycycline."
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
misc_feature 6791..7364
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(6958..6975)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 7185..7204
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 7380..8099
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 7380..8096
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(7425..7446)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(7686..7705)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 8033..8054
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 8235..8254
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
misc_feature 8542..9130
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(8595..8615)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(9013..9024)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 9205..9438
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(9557..9575)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 9643..9747
/gene="bla"
/label=AmpR promoter
CDS 9748..10608
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(9966..9985)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 10779..11367
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 11268..11287
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 11521..11538
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 11630..11649
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
protein_bind 11778..11799
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 11814..11844
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11852..11868
/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 11857..11879
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 11876..11892
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 11876..11892
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 11942..11961
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactaggatc
2221 ctttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
2281 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
2341 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
2401 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
2461 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg
2521 tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg
2581 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
2641 gaccgatcca gcctccgcgg ccccgaacta gtccagtgtg gtggaattct gcagatatca
2701 acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta
2761 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca
2821 ctatggcggc cgcattaggc accccaggct ttacacttta tgcttccggc tcgtataatg
2881 tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct aaaatggaga
2941 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
3001 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
3061 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
3121 ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
3181 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
3241 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
3301 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
3361 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
3421 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
3481 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
3541 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taaagatctg
3601 gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta
3661 taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca
3721 gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc
3781 aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa
3841 cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac
3901 ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa
3961 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
4021 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga
4081 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
4141 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
4201 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
4261 cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag tattatgtag
4321 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
4381 gttcagcttt cttgtacaaa gtggttgata tccagcacag tggcggccgc tcgagtctag
4441 agggcccgcg gttcgaatac ccatacgacg tcccagacta cgcttagtaa tgattaatta
4501 aactagaaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agtaacccac
4561 ccaagatctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcctcacgg
4621 cgagccgcgt aagatctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
4681 cctcacggcg agcctagtcg tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc
4741 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
4801 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
4861 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
4921 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
4981 atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg
5041 agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc
5101 ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc
5161 accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac
5221 ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca
5281 ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat
5341 gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag
5401 ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg
5461 caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg
5521 ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca
5581 cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc
5641 gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt
5701 ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta
5761 ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt
5821 ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt
5881 gaagcggccg ccatgctagt aaattgtccg ctaaattctg gccgtttttg gcttttttgt
5941 tagacggatc cataacttcg tataatgtat gctatacgaa gttatgccac catgtctaga
6001 ctggacaaga gcaaagtcat aaacggagct ctggaattac tcaatggtgt cggtatcgaa
6061 ggcctgacga caaggaaact cgctcaaaag ctgggagttg agcagcctac cctgtactgg
6121 cacgtgaaga acaagcgggc cctgctcgat gccctgccaa tcgagatgct ggacaggcat
6181 catacccact tctgccccct ggaaggcgag tcatggcaag actttctgcg gaacaacgcc
6241 aagtcatacc gctgtgctct cctctcacat cgcgacgggg ctaaagtgca tctcggcacc
6301 cgcccaacag agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa
6361 ggcttctccc tggagaacgc actgtacgct ctgtccgccg tgggccactt tacactgggc
6421 tgcgtattgg aggaacagga gcatcaagta gcaaaagagg aaagagagac acctaccacc
6481 gattctatgc ccccacttct gagacaagca attgagctgt tcgaccggca gggagccgaa
6541 cctgccttcc ttttcggcct ggaactaatc atatgtggcc tggagaaaca gctaaagtgc
6601 gaaagcggcg ggccgaccga cgcccttgac gattttgact tagacatgct cccagccgat
6661 gcccttgacg actttgacct tgatatgctg cctgctgacg ctcttgacga ttttgacctt
6721 gacatgctcc ccgggtaact aagtaaggat ccgcggccgc actagaggaa ttccgcccct
6781 ctcccccccc cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
6841 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
6901 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
6961 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
7021 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
7081 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
7141 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
7201 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
7261 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
7321 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttac cggtccacca
7381 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
7441 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
7501 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
7561 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
7621 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
7681 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
7741 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
7801 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
7861 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
7921 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
7981 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
8041 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag
8101 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
8161 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
8221 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
8281 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
8341 tcgccttctt gacgagttct tctgatgtac aagtaaagcg gccgcgactc tagatcataa
8401 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
8461 tgaacctgaa acataaaatg aatgcaattg ttgttgttta gtccctccca attcgatatc
8521 aagcttatcg cgatagatcc taatcaacct ctggattaca aaatttgtga aagattgact
8581 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
8641 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
8701 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
8761 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
8821 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
8881 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
8941 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
9001 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
9061 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
9121 gcctccccgc ctgagatcct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
9181 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
9241 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
9301 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
9361 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
9421 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
9481 caaagaaatg aatatcagag agtgagaggc ccgggttaat taaggaaagg gctagatcat
9541 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
9601 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
9661 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
9721 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
9781 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
9841 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
9901 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
9961 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
10021 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
10081 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
10141 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
10201 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
10261 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
10321 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
10381 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
10441 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
10501 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
10561 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
10621 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
10681 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
10741 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
10801 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
10861 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
10921 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
10981 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
11041 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
11101 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
11161 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
11221 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
11281 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
11341 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
11401 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
11461 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
11521 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
11581 cgttggccga ttcattaatg cagcaagctc atggctgact aatttttttt atttatgcag
11641 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
11701 gcctaggctt ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc
11761 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
11821 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
11881 aacagctatg acatgattac gaatttcaca aataaagcat ttttttcact gcattctagt
11941 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa
12001 gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg
12061 tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt
12121 atttgttaaa tatgtactac aaacttagta gt
//
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