| 质粒名称: | pFUGW ORANGE Tubb3-GFP KI _ mCherry-KASH |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 131508 |
| 存储实验室: | Harold MacGillavry |
|
载体骨架基本信息 |
|
| 载体名称: | pFUGW |
| 空载体大小: | 11480bp |
| 完整质粒大小: | 11472bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | gRNA and GFP donor |
| 物种来源: | M. musculus (mouse), R. norvegicus (rat) |
| 基因突变: | Tubb3 (a.k.a. 3200002H15Rik, M(beta)3, M(beta)6) |
| 启动子: | hUBC |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | 未知(未破坏) |
| 3’克隆位点: | 未知 (未破坏) |
| 5’测序引物 | GGCGAGTGTGTTTTGTGAAG |
| 3’测序引物 | GCAGCGTATCCACATAGCG |
|
文献引用方法 |
|
| 材料和方法部分: | pFUGW ORANGE Tubb3-GFP KI _ mCherry-KASH was a gift from Harold MacGillavry (Addgene plasmid # 131508 ; http://n2t.net/addgene:131508 ; RRID:Addgene_131508) |
| 参考文献部分: | ORANGE: A CRISPR/Cas9-based genome editing toolbox for epitope tagging of endogenous proteins in neurons. Willems J, de Jong APH, Scheefhals N, Mertens E, Catsburg LAE, Poorthuis RB, de Winter F, Verhaagen J, Meye FJ, MacGillavry HD. PLoS Biol. 2020 Apr 10;18(4):e3000665. doi: 10.1371/journal.pbio.3000665. 10.1371/journal.pbio.3000665 PubM |
买家导航
pFUGW ORANGE Tubb3-GFP KI _ mCherry-KASH载体质粒基本信息
pFUGW ORANGE Tubb3-GFP KI _ mCherry-KASH质粒图谱载体图谱和pFUGW ORANGE Tubb3-GFP KI _ mCherry-KASH载体序列质粒序列多克隆位点信息
pFUGW ORANGE Tubb3-GFP KI _ mCherry-KASH质粒载体简介
pFUGW ORANGE Tubb3-GFP KI _ mCherry-KASH质粒序列载体序列
LOCUS pFUGW ORANGE Tubb3-GFP KI _ mCherry-KASH 11472 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Endogenous tagging of beta-3 Tubulin: C-terminal (amino acid
position: STOP codon).
ACCESSION .
VERSION .
KEYWORDS pFUGW ORANGE Tubb3-GFP KI _ mCherry-KASH
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11472)
AUTHORS Willems J, de Jong APH, Scheefhals N, Mertens E, Catsburg LAE,
Poorthuis RB, de Winter F, Verhaagen J, Meye FJ, MacGillavry HD
TITLE ORANGE: A CRISPR/Cas9-based genome editing toolbox for epitope
tagging of endogenous proteins in neurons.
JOURNAL PLoS Biol. 2020 Apr 10;18(4):e3000665. doi:
10.1371/journal.pbio.3000665.
PUBMED 32275651
REFERENCE 2 (bases 1 to 11472)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..11472
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 277..394
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 436..676
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 436..456
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 607..626
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 704..779
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 906..1622
/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 906..1619
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(948..969)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1209..1228)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1556..1577
/label=EGFP-C
/note="EGFP, forward primer"
promoter 1741..2952
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 2789..2808
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
CDS 2999..3709
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MTVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYE
GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMN
FEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGAL
KGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYER
AEGRHSTGGMDELYK"
primer_bind complement(3147..3165)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3415..3434
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 4001..4589
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4054..4074)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4472..4483)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(4592..4608)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4593..4609)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 5114..5294
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(5320..5337)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5326..5550
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 5596..6024
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5683..5702)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5893..5914
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6033..6053)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 6038..6367
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6218..6353
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6280..6299
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 6415..6462
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 6481..6855
/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 6985..7106
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7022..7041)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7076..7095
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(7155..7171)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7155..7171)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7168..7190)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7179..7195
/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(7203..7233)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7248..7269
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7386..7403)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7557..8145)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7637..7656)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8316..9176)
/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 8939..8958
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9177..9281)
/gene="bla"
/label=AmpR promoter
primer_bind complement(9356..9375)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 9547..9926
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 9928..10126
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 10081..10101
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 10144..10324
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 10371..10496
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 10989..11222
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 11407..11451
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
ORIGIN
1 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
61 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
121 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
181 ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg
241 ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
301 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
361 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
421 tccagtttgg ttagtgaggg cctatttccc atgattcctt catatttgca tatacgatac
481 aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa
541 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
601 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
661 atcttgtgga aaggacgaaa caccgctgcg agcaacttca cttgttttag agctagaaat
721 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
781 tttttgtttt agagctagaa atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg
841 cgccaattct gcagacaaat ggctctagaa gcttgctgcg agcaacttca cttgggggat
901 caggcgtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
961 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
1021 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
1081 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
1141 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
1201 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
1261 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
1321 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
1381 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
1441 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
1501 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
1561 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
1621 aatgtgctgc gagcaacttc acttgggctc gaggatccta ggacgcgtcc taggacgcgt
1681 ccagaggtac ccgttacata acttacggta aatggcctaa cccgtgtcgg ctccagatct
1741 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg
1801 ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag
1861 cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag
1921 gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg
1981 aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat
2041 gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt
2101 cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct
2161 ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc
2221 caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg
2281 cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga
2341 ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgaggcctt
2401 cgctaatgcg ggaaagctct tattcgggtg agatgggctg gggcaccatc tggggaccct
2461 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
2521 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
2581 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg
2641 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
2701 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
2761 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
2821 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
2881 atatgtaatt ttcagtgtta gactagtaaa ttgtccgcta aattctggcc gtttttggct
2941 tttttgttag acgaagcttg ggctgcaggt cgactctaga ggatccccgg gtaccggtat
3001 gacggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
3061 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
3121 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
3181 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
3241 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
3301 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
3361 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
3421 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
3481 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
3541 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
3601 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
3661 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt ccggactcag
3721 atctcgagag gaggaggagg agacagacag caggatgccc cacctcgaca gccccggcag
3781 ctcccagccg agacgctcct tcctctcaag ggtgatcagg gcagcgctac cgttgcagct
3841 gcttctgctg ctgctgctgc tcctggcctg cctgctacct gcctctgaag atgactacag
3901 ctgcacccag gccaacaact ttgcccgatc cttctacccc atgctgcggt acaccaacgg
3961 gccacctccc acctaggaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
4021 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
4081 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
4141 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
4201 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
4261 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
4321 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
4381 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
4441 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
4501 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
4561 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgag acctagaaaa
4621 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
4681 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
4741 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
4801 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
4861 ctacttccct gattggcaga actacacacc agggccaggg atcagatatc cactgacctt
4921 tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg
4981 agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
5041 agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca
5101 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
5161 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
5221 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
5281 gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
5341 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
5401 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
5461 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
5521 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
5581 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
5641 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
5701 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
5761 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
5821 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
5881 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
5941 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
6001 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
6061 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
6121 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
6181 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
6241 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
6301 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
6361 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
6421 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
6481 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
6541 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
6601 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
6661 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
6721 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
6781 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
6841 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
6901 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
6961 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
7021 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
7081 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
7141 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
7201 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
7261 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
7321 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
7381 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
7441 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
7501 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
7561 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
7621 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
7681 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
7741 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
7801 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
7861 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
7921 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
7981 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
8041 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
8101 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
8161 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
8221 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
8281 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
8341 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
8401 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
8461 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
8521 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
8581 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
8641 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
8701 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
8761 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
8821 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
8881 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
8941 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
9001 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
9061 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
9121 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
9181 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
9241 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
9301 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
9361 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
9421 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
9481 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
9541 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
9601 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
9661 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
9721 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
9781 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
9841 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
9901 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
9961 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
10021 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
10081 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
10141 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
10201 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
10261 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
10321 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
10381 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
10441 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
10501 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
10561 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
10621 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
10681 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
10741 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
10801 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
10861 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
10921 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
10981 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
11041 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
11101 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
11161 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
11221 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
11281 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
11341 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
11401 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
11461 tttaacataa ca
//
