| 质粒名称: | TLCV2-APOBEC1-YTH |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 178949 |
| 存储实验室: | Kate Meyer |
|
载体骨架基本信息 |
|
| 载体名称: | lentiCRISPR v2 |
| 载体出品公司: | Feng Zhang lab |
| 空载体大小: | 11777bp |
| 完整质粒大小: | 13898bp |
| 载体类型: | 哺乳细胞表达, Lentiviral, Synthetic Biology ; Induc |
| 筛选标记: | Puromycin |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 30 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | APOBEC1-YTH-T2A-GFP |
| 物种来源: | H. sapiens (human), R. norvegicus (rat) |
| 插入大小: | 2121bp |
| 启动子: | tight TRE |
| 标签/融合蛋白: | HA (C terminal on insert) |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 |
LNCX
(Common Sequencing Primers)
|
|
文献引用方法 |
|
| 材料和方法部分: | TLCV2-APOBEC1-YTH was a gift from Kate Meyer (Addgene plasmid # 178949 ; http://n2t.net/addgene:178949 ; RRID:Addgene_178949) |
| 参考文献部分: | scDART-seq reveals distinct m(6)A signatures and mRNA methylation heterogeneity in single cells. Tegowski M, Flamand MN, Meyer KD. Mol Cell. 2022 Feb 17;82(4):868-878.e10. doi: 10.1016/j.molcel.2021.12.038. Epub 2022 Jan 25. 10.1016/j.molcel.2021.12.038 PubMed 35081365 |
买家导航
TLCV2-APOBEC1-YTH载体质粒基本信息
TLCV2-APOBEC1-YTH质粒图谱载体图谱和TLCV2-APOBEC1-YTH载体序列质粒序列多克隆位点信息
TLCV2-APOBEC1-YTH质粒载体简介
TLCV2-APOBEC1-YTH质粒序列载体序列
LOCUS TLCV2-APOBEC1-YTH 13898 bp DNA circular SYN 14-APR-2025
DEFINITION Lentiviral vector for dox-inducible expression of APOBEC1-YTH in
mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13898)
AUTHORS Tegowski M, Flamand MN, Meyer KD
TITLE scDART-seq reveals distinct m(6)A signatures and mRNA methylation
heterogeneity in single cells.
JOURNAL Mol Cell. 2022 Feb 17;82(4):868-878.e10. doi:
10.1016/j.molcel.2021.12.038. Epub 2022 Jan 25.
PUBMED 35081365
REFERENCE 2 (bases 1 to 13898)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.molcel.2021.12.038"; journalName: "Mol Cell"; date:
"2022-02-17- 17"; volume: "82"; issue: "4"; pages: "868-878.e10"
COMMENT Sequence Label: TLCV2-APOBEC1-YTH
FEATURES Location/Qualifiers
source 1..13898
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_RNA 238..313
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 350..664
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
protein_bind 358..376
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 394..412
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 429..447
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 465..483
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 501..519
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 536..554
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 572..590
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 637..661
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 696..1382
/codon_start=1
/product="cytidine deaminase (C to U editing enzyme) from
rat"
/label=APOBEC-1
/note="can use ssDNA as a substrate (Komor et al., 2016)"
/translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
LWATGLK"
CDS 2019..2045
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/note="This is a variant of the standard HA tag, which has
sequence YPYDVPDYA. Some anti-HA antibodies may not
recognize this variant."
/translation="YPYDVPDYA"
CDS 2019..2045
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 2046..2099
/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 join(2100..2102,2103..2816)
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
CDS join(2100..2102,2103..2813)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(2142..2163)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2403..2422)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2750..2771
/label=EGFP-C
/note="EGFP, forward primer"
promoter 2827..3038
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 3060..3656
/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(3060..3079)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3556..3576
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 3666..3722
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS join(3723..4340,4341..4346,4347..4382,4383..4385,4386..4421,
4422..4424,4425..4460,4461..4469)
/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 4490..5078
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4543..4563)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4961..4972)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5150..5383
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(5409..5426)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5415..5639
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 5685..6113
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5772..5791)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5982..6003
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6122..6142)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 6127..6456
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6307..6442
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6369..6388
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 6504..6551
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 6570..6944
/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 7074..7195
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7111..7130)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7165..7184
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(7244..7260)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7244..7260)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7257..7279)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7268..7284
/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(join(7292..7298,7299..7316,7317..7322))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7337..7358
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7475..7492)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7646..8234)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7726..7745)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(8405..9196,9197..9265))
/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 9028..9047
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9266..9370)
/gene="bla"
/label=AmpR promoter
primer_bind complement(9445..9464)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 9636..10015
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 10017..10215
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 10233..10413
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 10460..10585
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 11078..11311
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 11496..11540
/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 11838..11955
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 12006..12254
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 12006..12026
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 12177..12196
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
ORIGIN
1 tctgcatcat gtttaatacc atattcaagc ggtatttttc atgcaggatc aaattctgga
61 tttggattta caacattttt aaatgtttca tcttcaaatg catgctcttc aacctcaata
121 acgttatatc ctgattcacg taatattttt ggggtaaatt tagttcctgt tccattaact
181 gcgctaaaaa taatttttaa atctttttta gcttcttgct cttttttgta cgtctctgtt
241 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
301 accgagtcgg tgcttttttg aattcacttt ggccgcgaat cgatatgtcg agtttactcc
361 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgatgtc
421 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag
481 agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttatccct
541 atcagtgata gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg
601 aggtaggcgt gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat
661 cgcctggaga ataccggttc tagagcgctg ccaccatgag ctcagagact ggcccagtgg
721 ctgtggaccc cacattgaga cggcggatcg agccccatga gtttgaggta ttcttcgatc
781 cgagagagct ccgcaaggag acctgcctgc tttacgaaat taattggggg ggccggcact
841 ccatttggcg acatacatca cagaacacta acaagcacgt cgaagtcaac ttcatcgaga
901 agttcacgac agaaagatat ttctgtccga acacaaggtg cagcattacc tggtttctca
961 gctggagccc atgcggcgaa tgtagtaggg ccatcactga attcctgtca aggtatcccc
1021 acgtcactct gtttatttac atcgcaaggc tgtaccacca cgctgacccc cgcaatcgac
1081 aaggcctgcg ggatttgatc tcttcaggtg tgactatcca aattatgact gagcaggagt
1141 caggatactg ctggagaaac tttgtgaatt atagcccgag taatgaagcc cactggccta
1201 ggtatcccca tctgtgggta cgactgtacg ttcttgaact gtactgcatc atactgggcc
1261 tgcctccttg tctcaacatt ctgagaagga agcagccaca gctgacattc tttaccatcg
1321 ctcttcagtc ttgtcattac cagcgactgc ccccacacat tctctgggcc accgggttga
1381 aaagcggcag cgagactccc gggacctcag agtccgccac accagaaccc cacccagtgt
1441 tggagaagct tcggtccatt aataactata accccaaaga ttttgactgg aatctgaaac
1501 atggccgggt tttcatcatt aagagctact ctgaggacga tattcaccgt tccattaagt
1561 ataatatttg gtgcagcaca gagcatggta acaagagact ggatgctgct tatcgttcca
1621 tgaacgggaa aggccccgtt tacttacttt tcagtgtcaa cggcagtgga cacttctgtg
1681 gcgtggcaga aatgaaatct gctgtggact acaacacatg tgcaggtgtg tggtcccagg
1741 acaaatggaa gggtcgtttt gatgtcaggt ggatttttgt gaaggacgtt cccaatagcc
1801 aactgcgaca cattcgccta gagaacaacg agaataaacc agtgaccaac tctagggaca
1861 ctcaggaagt gcctctggaa aaggctaagc aggtgttgaa aattatagcc agctacaagc
1921 acaccacttc catttttgat gacttctcac actatgagaa acgccaagag gaagaagaaa
1981 gtgttaaaaa ggaacgtcaa ggtcgtggga aactcgagta cccctacgac gtgcccgact
2041 acgccgaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccag
2101 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
2161 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
2221 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
2281 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
2341 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
2401 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
2461 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
2521 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
2581 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
2641 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
2701 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
2761 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagcta
2821 gcatccgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
2881 attgatccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
2941 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
3001 acgttctttt tcgcaacggg tttgccgcca gaacacagac tgcgatcgca atgtacagta
3061 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
3121 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
3181 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
3241 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
3301 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
3361 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
3421 ccgcgtggtt cctggccacc gtcggagtct cgcccgacca ccagggcaag ggtctgggca
3481 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
3541 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
3601 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgccggtt
3661 ccggcgcaac aaacttctct ctgctgaaac aagccggaga tgtcgaagag aatcctggac
3721 cgatgtctag actggacaag agcaaagtca taaacggagc tctggaatta ctcaatggtg
3781 tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt gagcagccta
3841 ccctgtactg gcacgtgaag aacaagcggg ccctgctcga tgccctgcca atcgagatgc
3901 tggacaggca tcatacccac ttctgccccc tggaaggcga gtcatggcaa gactttctgc
3961 ggaacaacgc caagtcatac cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc
4021 atctcggcac ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc
4081 tgtgtcagca aggcttctcc ctggagaacg cactgtacgc tctgtccgcc gtgggccact
4141 ttacactggg ctgcgtattg gaggaacagg agcatcaagt agcaaaagag gaaagagaga
4201 cacctaccac cgattctatg cccccacttc tgagacaagc aattgagctg ttcgaccggc
4261 agggagccga acctgccttc cttttcggcc tggaactaat catatgtggc ctggagaaac
4321 agctaaagtg cgaaagcggc gggccgaccg acgcccttga cgattttgac ttagacatgc
4381 tcccagccga tgcccttgac gactttgacc ttgatatgct gcctgctgac gctcttgacg
4441 attttgacct tgacatgctc cccgggtaac tcgagcgcgt taagtcgaca atcaacctct
4501 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
4561 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
4621 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
4681 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
4741 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
4801 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
4861 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
4921 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
4981 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
5041 tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact
5101 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
5161 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
5221 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
5281 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
5341 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg
5401 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
5461 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
5521 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
5581 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
5641 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
5701 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
5761 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
5821 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
5881 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
5941 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
6001 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
6061 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
6121 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
6181 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
6241 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
6301 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
6361 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
6421 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
6481 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
6541 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
6601 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
6661 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
6721 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
6781 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
6841 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
6901 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt
6961 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
7021 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
7081 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
7141 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
7201 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
7261 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
7321 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
7381 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
7441 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
7501 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
7561 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
7621 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
7681 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
7741 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
7801 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
7861 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
7921 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
7981 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
8041 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
8101 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
8161 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
8221 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
8281 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
8341 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
8401 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
8461 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
8521 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
8581 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
8641 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
8701 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
8761 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
8821 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
8881 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
8941 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
9001 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
9061 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
9121 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
9181 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
9241 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
9301 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
9361 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
9421 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
9481 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
9541 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
9601 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
9661 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
9721 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
9781 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
9841 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
9901 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
9961 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
10021 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
10081 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
10141 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
10201 ggtctatata agcagcgcgt tttgcctgta ctgggtctct ctggttagac cagatctgag
10261 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
10321 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
10381 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
10441 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
10501 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
10561 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
10621 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
10681 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
10741 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
10801 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
10861 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
10921 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
10981 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
11041 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
11101 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
11161 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
11221 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
11281 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
11341 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
11401 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
11461 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
11521 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
11581 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
11641 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
11701 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag
11761 acagatccat tcgattagtg aacggatcgg cactgcgtgc gccaattctg cagacaaatg
11821 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
11881 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
11941 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gttaattaag
12001 gtaccgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
12061 gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
12121 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
12181 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
12241 aaggacgaaa caccggagac ggttgtaaat gagcacacaa aatacacatg ctaaaatatt
12301 atattctatg acctttataa aatcaaccaa aatcttcttt ttaataactt tagtatcaat
12361 aattagaatt tttatgttcc tttttgcaaa cttttaataa aaatgagcaa aataaaaaaa
12421 cgctagtttt agtaactcgc gttgttttct tcacctttaa taatagctac tccaccactt
12481 gttcctaagc ggtcagctcc tgcttcaatc attttttgag catcttcaaa tgttctaact
12541 ccaccagctg ctttaactaa agcattgtct ttaacaactg acttcattag tttaacatct
12601 tcaaatgttg cacctgattt tgaaaatcct gttgatgttt taacaaattc taatccagct
12661 tcaacagcta tttcacaagc tttcatgatt tcttcttttg ttaataaaca attttccata
12721 atacatttaa caacatgtga tccagctgct ttttttacag ctttcatgtc ttctaaaact
12781 aattcataat ttttgtcttt taatgcacca atatttaata ccatatcaat ttctgttgca
12841 ccatctttaa ttgcttcaga aacttcgaat gcttttgtag ctgttgtgca tgcacctaga
12901 ggaaaaccta caacatttgt tattcctaca tttgtgcctt ttaataattc tttacaatag
12961 cttgttcaat atgaattaac acaaactgtt gcaaaatcaa attcaattgc ttcatcacat
13021 aattgtttaa tttcagcttt cgtagcatct tgttttaata atgtgtgatc tatatatttg
13081 tttagtttca ttttttctcc tatatattca tttttaattt taattcttta ataatttcgt
13141 ctactttaac tttagcgttt tgaacagatt caccaacacc tataaaataa atttttagtt
13201 taggttcagt tccacttggg cgaacagcaa atcatgactt atcttctaaa taaaatttta
13261 gtaagtcttg tcctggcata ttatacattc catcgatgta gtcttcaaca ttaacaactt
13321 taagtccagc aatttgagtt aagggtgttg ctctcaatga tttcattaat ggttcaattt
13381 ttaatttctt ttcttctggt ttaaaattca agtttaaagt gaaagtgtaa tatgcaccca
13441 tttctttaaa taaatcttct aaatagtcta ctaatgtttt attttgtttt ttataaaatc
13501 aagcagcctc tgctattaat atagaagctt gtattccatc tttatctcta gctgagtcat
13561 caattacata tccataactt tcttcataag caaaaacaaa atttaatccg ttatcttctt
13621 ctttagcaat ttctctaccc attcatttaa atccagttaa agtttttaca atattaactc
13681 catatttttc atgagcgatt ctatcaccca aatcacttgt tacaaaactt gaatatagag
13741 ccggattttt tggaatgcta tttaagcgtt ttagatttga taattttcaa tcaattaaaa
13801 ttggtcctgt ttgatttcca tctaatctta caaaatgacc atcatgtttt attgccattc
13861 caaatctgtc agcatctggg tcattcataa taataata
//
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