| 质粒名称: | TLCV2-APOBEC1-YTHmut |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 178950 |
| 存储实验室: | Kate Meyer |
|
载体骨架基本信息 |
|
| 载体名称: | TLCV2 (lentiCRISPR v2) |
| 载体出品公司: | Adam Karpf (Feng Zhang) |
| 空载体大小: | 11775bp |
| 完整质粒大小: | 13823bp |
| 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-YTHmut-T2A-GFP |
| 物种来源: | H. sapiens (human), R. norvegicus (rat) |
| 插入大小: | 2046bp |
| 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 |
|
文献引用方法 |
|
| 材料和方法部分: | TLCV2-APOBEC1-YTHmut was a gift from Kate Meyer (Addgene plasmid # 178950 ; http://n2t.net/addgene:178950 ; RRID:Addgene_178950) |
| 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-YTHmut载体质粒基本信息
TLCV2-APOBEC1-YTHmut质粒图谱载体图谱和TLCV2-APOBEC1-YTHmut载体序列质粒序列多克隆位点信息
TLCV2-APOBEC1-YTHmut质粒载体简介
TLCV2-APOBEC1-YTHmut质粒序列载体序列
LOCUS TLCV2-APOBEC1-YTHmut 13823 bp DNA circular SYN 14-APR-2025
DEFINITION Lentiviral vector for dox-inducible expression of APOBEC1-YTHmut in
mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13823)
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 13823)
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-YTHmut
FEATURES Location/Qualifiers
source 1..13823
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_RNA 291..366
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 403..717
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
protein_bind 411..429
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 447..465
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 482..500
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 518..536
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 554..572
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 589..607
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 625..643
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 690..714
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 749..1435
/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 1997..2023
/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 1997..2023
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 2024..2077
/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(2078..2080,2081..2794)
/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(2078..2080,2081..2791)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(2120..2141)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2381..2400)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2728..2749
/label=EGFP-C
/note="EGFP, forward primer"
promoter 2805..3016
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 3038..3634
/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(3038..3057)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3534..3554
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 3644..3700
/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(3701..4318,4319..4324,4325..4360,4361..4363,4364..4399,
4400..4402,4403..4438,4439..4447)
/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 4468..5056
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4521..4541)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4939..4950)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5128..5361
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(5387..5404)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5393..5617
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 5663..6091
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5750..5769)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5960..5981
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6100..6120)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 6105..6434
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6285..6420
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6347..6366
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 6482..6529
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 6548..6922
/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 7052..7173
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7089..7108)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7143..7162
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(7222..7238)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7222..7238)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7235..7257)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7246..7262
/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(7270..7276,7277..7294,7295..7300))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7315..7336
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7453..7470)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7624..8212)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7704..7723)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(8383..9174,9175..9243))
/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 9006..9025
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9244..9348)
/gene="bla"
/label=AmpR promoter
primer_bind complement(9423..9442)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 9614..9993
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 9995..10193
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 10211..10391
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 10438..10563
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 11056..11289
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 11474..11518
/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 11816..11933
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 11984..12232
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 11984..12004
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 12155..12174
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
ORIGIN
1 gttttattgc cattccaaat ctgtcagcat ctgggtcatt cataataata atatctgcat
61 catgtttaat accatattca agcggtattt ttcatgcagg atcaaattct ggatttggat
121 ttacaacatt tttaaatgtt tcatcttcaa atgcatgctc ttcaacctca ataacgttat
181 atcctgattc acgtaatatt tttggggtaa atttagttcc tgttccatta actgcgctaa
241 aaataatttt taaatctttt ttagcttctt gctctttttt gtacgtctct gttttagagc
301 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
361 cggtgctttt ttgaattcac tttggccgcg aatcgatatg tcgagtttac tccctatcag
421 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgat gtcgagttta
481 ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt
541 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttatc cctatcagtg
601 atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgaggtagg
661 cgtgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
721 agaataccgg ttctagagcg ctgccaccat gagctcagag actggcccag tggctgtgga
781 ccccacattg agacggcgga tcgagcccca tgagtttgag gtattcttcg atccgagaga
841 gctccgcaag gagacctgcc tgctttacga aattaattgg gggggccggc actccatttg
901 gcgacataca tcacagaaca ctaacaagca cgtcgaagtc aacttcatcg agaagttcac
961 gacagaaaga tatttctgtc cgaacacaag gtgcagcatt acctggtttc tcagctggag
1021 cccatgcggc gaatgtagta gggccatcac tgaattcctg tcaaggtatc cccacgtcac
1081 tctgtttatt tacatcgcaa ggctgtacca ccacgctgac ccccgcaatc gacaaggcct
1141 gcgggatttg atctcttcag gtgtgactat ccaaattatg actgagcagg agtcaggata
1201 ctgctggaga aactttgtga attatagccc gagtaatgaa gcccactggc ctaggtatcc
1261 ccatctgtgg gtacgactgt acgttcttga actgtactgc atcatactgg gcctgcctcc
1321 ttgtctcaac attctgagaa ggaagcagcc acagctgaca ttctttacca tcgctcttca
1381 gtcttgtcat taccagcgac tgcccccaca cattctctgg gccaccgggt tgaaaagcgg
1441 cagcgagact cccgggacct cagagtccgc cacaccagaa ggccgggttt tcatcattaa
1501 gagctactct gaggacgata ttcaccgttc cattaagtat aatatttggt gcagcacaga
1561 gcatggtaac aagagactgg atgctgctta tcgttccatg aacgggaaag gccccgttta
1621 cttacttttc agtgtcaacg gcagtggaca cttctgtggc gtggcagaaa tgaaatctgc
1681 tgtggactac aacacatgtg caggtgtgtg gtcccaggac aaatggaagg gtcgttttga
1741 tgtcaggtgg atttttgtga aggacgttcc caatagccaa ctgcgacaca ttcgcctaga
1801 gaacaacgag aataaaccag tgaccaactc tagggacact caggaagtgc ctctggaaaa
1861 ggctaagcag gtgttgaaaa ttatagccag ctacaagcac accacttcca tttttgatga
1921 cttctcacac tatgagaaac gccaagagga agaagaaagt gttaaaaagg aacgtcaagg
1981 tcgtgggaaa ctcgagtacc cctacgacgt gcccgactac gccgagggca gaggaagtct
2041 gctaacatgc ggtgacgtcg aggagaatcc tggcccagtg agcaagggcg aggagctgtt
2101 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
2161 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
2221 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
2281 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
2341 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
2401 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
2461 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
2521 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
2581 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
2641 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
2701 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
2761 gatcactctc ggcatggacg agctgtacaa gtaagctagc atccgggcag agcgcacatc
2821 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgatccggtg cctagagaag
2881 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
2941 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
3001 tgccgccaga acacagactg cgatcgcaat gtacagtatg accgagtaca agcccacggt
3061 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
3121 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
3181 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
3241 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
3301 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
3361 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
3421 cggagtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
3481 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
3541 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
3601 gcgcacctgg tgcatgaccc gcaagcccgg tgccggttcc ggcgcaacaa acttctctct
3661 gctgaaacaa gccggagatg tcgaagagaa tcctggaccg atgtctagac tggacaagag
3721 caaagtcata aacggagctc tggaattact caatggtgtc ggtatcgaag gcctgacgac
3781 aaggaaactc gctcaaaagc tgggagttga gcagcctacc ctgtactggc acgtgaagaa
3841 caagcgggcc ctgctcgatg ccctgccaat cgagatgctg gacaggcatc atacccactt
3901 ctgccccctg gaaggcgagt catggcaaga ctttctgcgg aacaacgcca agtcataccg
3961 ctgtgctctc ctctcacatc gcgacggggc taaagtgcat ctcggcaccc gcccaacaga
4021 gaaacagtac gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag gcttctccct
4081 ggagaacgca ctgtacgctc tgtccgccgt gggccacttt acactgggct gcgtattgga
4141 ggaacaggag catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc
4201 cccacttctg agacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct
4261 tttcggcctg gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg
4321 gccgaccgac gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga
4381 ctttgacctt gatatgctgc ctgctgacgc tcttgacgat tttgaccttg acatgctccc
4441 cgggtaactc gagcgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga
4501 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
4561 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
4621 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
4681 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
4741 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
4801 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
4861 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
4921 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
4981 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
5041 tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta
5101 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
5161 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
5221 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
5281 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
5341 cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct
5401 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
5461 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
5521 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
5581 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
5641 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
5701 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
5761 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
5821 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
5881 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
5941 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
6001 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
6061 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
6121 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
6181 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
6241 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
6301 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
6361 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
6421 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
6481 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
6541 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
6601 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
6661 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
6721 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
6781 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
6841 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
6901 cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta
6961 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
7021 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
7081 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
7141 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
7201 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
7261 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
7321 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
7381 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
7441 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
7501 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
7561 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
7621 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
7681 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
7741 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
7801 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
7861 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
7921 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
7981 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
8041 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
8101 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
8161 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
8221 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
8281 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
8341 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
8401 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
8461 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
8521 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
8581 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
8641 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
8701 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
8761 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
8821 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
8881 acataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
8941 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
9001 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
9061 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
9121 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
9181 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
9241 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
9301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
9361 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
9421 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
9481 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
9541 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
9601 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
9661 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
9721 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
9781 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
9841 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
9901 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
9961 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
10021 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
10081 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
10141 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt
10201 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
10261 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
10321 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
10381 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
10441 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
10501 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
10561 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
10621 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
10681 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
10741 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
10801 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
10861 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
10921 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
10981 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
11041 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
11101 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
11161 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
11221 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
11281 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
11341 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
11401 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
11461 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
11521 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
11581 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
11641 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
11701 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
11761 cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta
11821 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
11881 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
11941 attacaggga cagcagagat ccagtttggt taattaaggt accgagggcc tatttcccat
12001 gattccttca tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt
12061 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
12121 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
12181 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccggagacgg
12241 ttgtaaatga gcacacaaaa tacacatgct aaaatattat attctatgac ctttataaaa
12301 tcaaccaaaa tcttcttttt aataacttta gtatcaataa ttagaatttt tatgttcctt
12361 tttgcaaact tttaataaaa atgagcaaaa taaaaaaacg ctagttttag taactcgcgt
12421 tgttttcttc acctttaata atagctactc caccacttgt tcctaagcgg tcagctcctg
12481 cttcaatcat tttttgagca tcttcaaatg ttctaactcc accagctgct ttaactaaag
12541 cattgtcttt aacaactgac ttcattagtt taacatcttc aaatgttgca cctgattttg
12601 aaaatcctgt tgatgtttta acaaattcta atccagcttc aacagctatt tcacaagctt
12661 tcatgatttc ttcttttgtt aataaacaat tttccataat acatttaaca acatgtgatc
12721 cagctgcttt ttttacagct ttcatgtctt ctaaaactaa ttcataattt ttgtctttta
12781 atgcaccaat atttaatacc atatcaattt ctgttgcacc atctttaatt gcttcagaaa
12841 cttcgaatgc ttttgtagct gttgtgcatg cacctagagg aaaacctaca acatttgtta
12901 ttcctacatt tgtgcctttt aataattctt tacaatagct tgttcaatat gaattaacac
12961 aaactgttgc aaaatcaaat tcaattgctt catcacataa ttgtttaatt tcagctttcg
13021 tagcatcttg ttttaataat gtgtgatcta tatatttgtt tagtttcatt ttttctccta
13081 tatattcatt tttaatttta attctttaat aatttcgtct actttaactt tagcgttttg
13141 aacagattca ccaacaccta taaaataaat ttttagttta ggttcagttc cacttgggcg
13201 aacagcaaat catgacttat cttctaaata aaattttagt aagtcttgtc ctggcatatt
13261 atacattcca tcgatgtagt cttcaacatt aacaacttta agtccagcaa tttgagttaa
13321 gggtgttgct ctcaatgatt tcattaatgg ttcaattttt aatttctttt cttctggttt
13381 aaaattcaag tttaaagtga aagtgtaata tgcacccatt tctttaaata aatcttctaa
13441 atagtctact aatgttttat tttgtttttt ataaaatcaa gcagcctctg ctattaatat
13501 agaagcttgt attccatctt tatctctagc tgagtcatca attacatatc cataactttc
13561 ttcataagca aaaacaaaat ttaatccgtt atcttcttct ttagcaattt ctctacccat
13621 tcatttaaat ccagttaaag tttttacaat attaactcca tatttttcat gagcgattct
13681 atcacccaaa tcacttgtta caaaacttga atatagagcc ggattttttg gaatgctatt
13741 taagcgtttt agatttgata attttcaatc aattaaaatt ggtcctgttt gatttccatc
13801 taatcttaca aaatgaccat cat
//
