| 质粒名称: | pINDUCER11 (miR-RUG) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 44363 |
| 存储实验室: | Thomas Westbrook |
|
载体骨架基本信息 |
|
| 载体名称: | GIPZ |
| 载体出品公司: | Open Biosystems |
| 载体抗性: | 氨苄西林 |
| 空载体大小: | 11688 bp |
| 完整质粒大小: | 14699 bp |
| 载体修饰: | 首先用Xbai和Paci消化pinducer10，通过puromycin抗性基因去除RTTA，制备了pinducer11（mir rug）。Xbai和Noti Digest从pindmir3luc-mir中删除了rttaires段。用引物5′-taatacatgt-atcgatccatggtgaggcaaggc和5′-gatcttaattaa-ttcgatccatggccatggccgc从mscv-n-egfp-entr扩增egfp。PCR产物经PCII和PACI消化。然后将载体、rtta-ires和egfp连接起来并验证序列。 |
| 载体类型: | 慢病毒载体 |
| 筛选标记: | EGFP; RFP |
| 高拷贝/低拷贝: | 高拷贝 |
| 宿生菌： | Stbl3 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | PheS Gly294 |
| GRNA/SHRNA序列： | mir30 |
| 物种来源: | 浮游盐杆菌P. haloplanktis |
|
文献引用方法 |
|
| 材料和方法部分: | pINDUCER11 (miR-RUG) was a gift from Thomas Westbrook (Addgene plasmid # 44363 ; http://n2t.net/addgene:44363 ; RRID:Addgene_44363) |
| 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
载体基本信息
载体图谱质粒图谱和多克隆位点信息
质粒图谱.png)
载体简介
载体序列
LOCUS pINDUCER11 (miR-RUG) 14701 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Inducible lentiviral gene silencing vector. Insert PheSGly294, (XhoI
to EcoR1; 1.4kb), can be replaced w mir30 based hairpin of interest.
ACCESSION .
VERSION .
KEYWORDS pINDUCER11 (miR-RUG)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14701)
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 14701)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14701
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 500..733
/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 1260..1377
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(1442..1816)
/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"
regulatory 1813..1822
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
promoter complement(1826..1844)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(1836..1883)
/label=EM7 promoter
/note="synthetic bacterial promoter "
protein_bind 1953..2223
/label=tetracycline response element
/bound_moiety="tetracycline repressor TetR"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 2256..2294
/label=minimal CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2291..2315
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 2391..3086
/codon_start=1
/product="red fluorescent protein from Entacmaea
quadricolor"
/label=TurboRFP
/note="mammalian codon-optimized"
/translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
DLPSKLGHR"
ncRNA 3137..3230
/label=5' miR-30a
/note="sequence upstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
/ncRNA_class="miRNA"
primer_bind 3230..3246
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 3231..3247
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 3268..3285
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 3286..3309
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
regulatory 3319..3328
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4604..4624
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
ncRNA 4648..4760
/label=3' miR-30a
/note="sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
/ncRNA_class="miRNA"
promoter 4811..6022
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 5859..5878
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
protein_bind complement(6029..6062)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS 6069..6815
/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 6864..7436
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(7031..7048)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 7258..7277
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 7454..8173
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(7499..7520)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(7760..7779)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 8107..8128
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 8195..8783
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(8248..8268)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(8666..8677)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 8990..9223
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(9324..9341)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 9330..9554
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 9600..10028
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(9687..9706)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 9897..9918
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(10037..10057)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 10042..10371
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 10222..10357
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 10284..10303
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 11194..11315
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(11364..11380)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(11364..11380)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(11377..11399)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 11388..11404
/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(11412..11442)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11457..11478
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(11595..11612)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(11766..12354)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(11846..11865)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(12525..13385)
/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 13148..13167
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(13386..13490)
/gene="bla"
/label=AmpR promoter
polyA_signal 13538..13672
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
LTR 13840..14473
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 14520..14645
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
ORIGIN
1 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
61 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
121 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
181 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
241 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
301 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca
361 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
421 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
481 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
541 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
601 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
661 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
721 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
781 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
841 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
901 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
961 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
1021 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
1081 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
1141 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
1201 tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat
1261 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
1321 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
1381 gtttattaca gggacagcag agatccagtt tggttagtac cgggcccgct ctagtccgga
1441 atcagtcctg ctcctcggcc acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga
1501 actcccgccc ccacggctgc tcgccgatct cggtcatggc cggcccggag gcgtcccgga
1561 agttcgtgga cacgacctcc gaccactcgg cgtacagctc gtccaggccg cgcacccaca
1621 cccaggccag ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg aacagggtca
1681 cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc
1741 ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac
1801 tggtcaactt ggccatggtg gccctcctat agtgagtcgt attatactat gccgatatac
1861 tatgccgatg attaattgtc aacacgtgct gcaggtccga ggttctaggc tagcttaaag
1921 gaaccaattc agtcgactgg atcctttacc actccctatc agtgatagag aaaagtgaaa
1981 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
2041 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
2101 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
2161 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
2221 agaaaagtga aagtcgagct cggtacccgg gtcgaggtag gcgtgtacgg tgggaggcct
2281 atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
2341 ttgacctcca tagaagacac cgggaccgat ccagcctacc ggtcgccacc atgagcgagc
2401 tgatcaagga gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact
2461 tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc atgaagatca
2521 aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt
2581 acggcagcaa agccttcatc aaccacaccc agggcatccc cgacttcttt aagcagtcct
2641 tccctgaggg cttcacatgg gagagaatca ccacatacga agacgggggc gtgctgaccg
2701 ctacccagga caccagcttc cagaacggct gcatcatcta caacgtcaag atcaacgggg
2761 tgaacttccc atccaacggc cctgtgatgc agaagaaaac acgcggctgg gaggccaaca
2821 ccgagatgct gtaccccgct gacggcggcc tgagaggcca cagccagatg gccctgaagc
2881 tcgtgggcgg gggctacctg cactgctcct tcaagaccac atacagatcc aagaaacccg
2941 ctaagaacct caagatgccc ggcttccact tcgtggacca cagactggaa agaatcaagg
3001 aggccgacaa agagacctac gtcgagcagc acgagatggc tgtggccaag tactgcgacc
3061 tccctagcaa actggggcac agatgatgcg gccgcaagcc ttgttaagtg ctcgcttcgg
3121 cagcacatat actatgttga atgaggcttc agtactttac agaatcgttg cctgcacatc
3181 ttggaaacac ttgctgggat tacttcttca ggttaaccca acagaaggct cgaggtcgac
3241 ggtatcgata agcttgacgt ccatatgctg gtaccccgtg gaagtctgga agttctcttc
3301 caggggcccg aaggagacgc caccatgggt ctagataggg ataacagggt aatggtaccg
3361 gatcgatccc ccaacttcca ggcccgccat agtgatttga tttgccagcg catcgctatc
3421 aatcgccggg ttcacccatt cgcgtaacca cagttcactg aatttcataa tctattcctg
3481 ccttatttaa actgtttgag gaaacgcaga tcgttttcga agaatgaacg caggtcggtg
3541 acgccgtaac gcaacatagt cagacgctcc atccccatcc cgaagccgaa accagagtaa
3601 acttccgggt cgatgccaac gttacgcaac acgttcggat gcaccatccc gcagcccagc
3661 acttccagcc atttaccgtt tttacccatg acgtccactt ctgcagaagg ttcggtaaac
3721 gggaagtagg aaggacggaa gcgaatctgc aaatcttcct caaagaagtt acgcaggaag
3781 tcgtgcagcg tgcctttcag gttggtaaag ctgatgttgg tatcaacaat cagaccttcc
3841 atctgatgga acatcggcgt gtgagtctgg tcgtagtcgt tacgataaac acggccaggc
3901 gcgatgatac gaatcggtgg ctgctgggct ttcatggtgc ggatctgtac gccagaggtc
3961 tgggtacgca gcaggcgggt agtgtcaaac cagaaagtgt cgtggtcagc gcgcgccggg
4021 tggtgaccag gaatgttcag agcatcgaag ttatgataat cgtcttcgat ttccggcccg
4081 gttgccacgg taaagccaag ctcaccgaag aaactttcga tacggtcgat ggtacgggta
4141 accggatgca gaccgccgtt ttcaatgcga cgacctggca gagagacatc aatcgtttcc
4201 gccgccagac gcgcattcag tgcagcgctt tccagttccg ctttacgcgc attcagcgcc
4261 tgctgaacct gctctttcgc ttcgttgata accgcaccag ctgccggacg ctcttctggc
4321 ggcagctcac gcagggtcgt catctgaagg gttaagtgcc cttttttacc caaatattcg
4381 acgcgcacat tatctaacgc ggcaacatct gacgcctggc taatggccgc cttcgcactg
4441 gcaaccagtt ctgcgagatg tgacatggtt ttcctcattg tgtcagtggt gacactggtt
4501 cgttggactt agagcctatc ccatcaggct attttacttg ccattttggt ccccgatacc
4561 gtcgaccgcg gattaccctg ttatccctag atctcacaat tctgagaatt tgtattttca
4621 gggtgatctc cgtggatcta ttacgaattc aaggggctac tttaggagca attatcttgt
4681 ttactaaaac tgaatacctt gctatctctt tgatacattt ttacaaagct gaattaaaat
4741 ggtataaatt aaatcacttt tttcaattgg aagactaatg cgtttaaaca cgcggcgacg
4801 cgtaagatct ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg
4861 gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg
4921 ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag
4981 gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga
5041 acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt
5101 gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg
5161 ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg
5221 ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg
5281 gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt
5341 ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg
5401 cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc
5461 ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg gggcaccatc
5521 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
5581 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
5641 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc
5701 ggtaggtgtg tggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
5761 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
5821 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
5881 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
5941 atttgggtca atatgtaatt ttcagtgtta gactagtaaa ttgtccgcta aattctggcc
6001 gtttttggct tttttgttag acggatccat aacttcgtat aatgtatgct atacgaagtt
6061 atgccaccat gtctagactg gacaagagca aagtcataaa cggagctctg gaattactca
6121 atggtgtcgg tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc
6181 agcctaccct gtactggcac gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg
6241 agatgctgga caggcatcat acccacttct gccccctgga aggcgagtca tggcaagact
6301 ttctgcggaa caacgccaag tcataccgct gtgctctcct ctcacatcgc gacggggcta
6361 aagtgcatct cggcacccgc ccaacagaga aacagtacga aaccctggaa aatcagctcg
6421 cgttcctgtg tcagcaaggc ttctccctgg agaacgcact gtacgctctg tccgccgtgg
6481 gccactttac actgggctgc gtattggagg aacaggagca tcaagtagca aaagaggaaa
6541 gagagacacc taccaccgat tctatgcccc cacttctgag acaagcaatt gagctgttcg
6601 accggcaggg agccgaacct gccttccttt tcggcctgga actaatcata tgtggcctgg
6661 agaaacagct aaagtgcgaa agcggcgggc cgaccgacgc ccttgacgat tttgacttag
6721 acatgctccc agccgatgcc cttgacgact ttgaccttga tatgctgcct gctgacgctc
6781 ttgacgattt tgaccttgac atgctccccg ggtaaataac ttcgtatagc atacattata
6841 cgaagtatgg cgcgccaatt ccgcccctct ccctcccccc cccctaacgt tactggccga
6901 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
6961 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
7021 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
7081 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
7141 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
7201 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
7261 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
7321 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
7381 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg ataataccat
7441 gtatcgatcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
7501 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
7561 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
7621 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
7681 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
7741 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
7801 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
7861 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
7921 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
7981 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
8041 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
8101 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
8161 gctgtacaag taattaatta agcgcgtctg gaacaatcaa cctctggatt acaaaatttg
8221 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
8281 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
8341 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
8401 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
8461 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
8521 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
8581 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
8641 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
8701 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
8761 ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat
8821 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
8881 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
8941 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta
9001 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
9061 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
9121 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
9181 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
9241 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga
9301 cattgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
9361 ttgccccttc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
9421 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
9481 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
9541 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
9601 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
9661 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
9721 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
9781 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
9841 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
9901 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
9961 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
10021 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
10081 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
10141 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
10201 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
10261 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
10321 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
10381 ggagcttgta tatccatttt cggatctgat cagcacgtga tgaaaaagcc tgaactcacc
10441 gcgacgtgac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt
10501 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc
10561 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc
10621 gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc
10681 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc
10741 tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg
10801 gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc
10861 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac
10921 gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg
10981 gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc
11041 actcgtccga gggcaaagga atagcacgtg ctacgagatt tcgattccac cgccgccttc
11101 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
11161 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
11221 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
11281 agttgtggtt tgtccaaact aatcaatgta tcttatcatg tctgtatacc gtcgacctct
11341 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
11401 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
11461 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
11521 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
11581 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
11641 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
11701 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
11761 gcgtttttcc ataggctccg cccccctgat gagcatcaca aaaatcgacg ctcaagtcag
11821 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
11881 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
11941 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
12001 cgctccaagc tgggctgtgt gcactaaccc cccgttcagc ccgaccgctg cgccttatcc
12061 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
12121 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
12181 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
12241 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
12301 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
12361 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
12421 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
12481 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
12541 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
12601 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
12661 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
12721 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
12781 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
12841 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
12901 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
12961 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
13021 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
13081 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
13141 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
13201 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
13261 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
13321 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
13381 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
13441 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
13501 cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac ttgtttattg cagcttataa
13561 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
13621 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat
13681 aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa
13741 ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag
13801 aaattgtatt tgttaaatat gtactacaaa cttagtagtt ggaagggcta attcactccc
13861 aaagaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt
13921 agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca
13981 agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct
14041 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga
14101 ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca
14161 agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc
14221 gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt
14281 tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
14341 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
14401 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
14461 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
14521 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
14581 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
14641 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttarggagag
14701 c
//
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