| 质粒名称: | pKLV2-U6gRNA5(Empty)-PGKBFP2AGFP-W |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 67979 |
| 存储实验室: | Kosuke Yusa |
|
载体骨架基本信息 |
|
| 载体名称: | pKLV2 lentiviral vector |
| 载体出品公司: | Clontech |
| 空载体大小: | 5783bp |
| 完整质粒大小: | 8767bp |
| 载体类型: | Lentiviral |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | U6gRNA cassette, PGKBFP2AGFP cassette, WPRE |
| 物种来源: | H. sapiens (human), M. musculus (mouse) |
| 插入大小: | 3022bp |
| 基因突变: | Deleted BbsI site within WPRE |
| 启动子: | human U6 promoter, mouse PGK promoter |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | MluI (not destroyed) |
| 3’克隆位点: | KpnI (not destroyed) |
| 5’测序引物 | CATAATAGCAACAGACATAC |
|
文献引用方法 |
|
| 材料和方法部分: | pKLV2-U6gRNA5(Empty)-PGKBFP2AGFP-W was a gift from Kosuke Yusa (Addgene plasmid # 67979 ; http://n2t.net/addgene:67979 ; RRID:Addgene_67979) |
| 参考文献部分: | A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and Therapeutic Targets in Acute Myeloid Leukemia. Tzelepis K, Koike-Yusa H, De Braekeleer E, Li Y, Metzakopian E, Dovey OM, Mupo A, Grinkevich V, Li M, Mazan M, Gozdecka M, Ohnishi S, Cooper J, Patel M, McKerrell T, Chen B, Domingues AF, Gallipoli P, Teichmann S, Ponstingl H, McDermott U, Saez-Rodriguez J, Huntly BJ, Iorio F, Pina C, Vassiliou GS, Yusa K. Cell Rep. 2016 Oct 18;17(4):1193-1205. doi: 10.1016/j.celrep.2016.09.079. |
买家导航
pKLV2-U6gRNA5(Empty)-PGKBFP2AGFP-W载体质粒基本信息
pKLV2-U6gRNA5(Empty)-PGKBFP2AGFP-W质粒图谱载体图谱和 pKLV2-U6gRNA5(Empty)-PGKBFP2AGFP-W载体序列质粒序列多克隆位点信息
-PGKBFP2AGFP-W质粒图谱.png)
pKLV2-U6gRNA5(Empty)-PGKBFP2AGFP-W质粒载体简介
pKLV2-U6gRNA5(Empty)-PGKBFP2AGFP-W质粒序列载体序列
LOCUS pKLV2-U6gRNA5(Empty)-PGKBFP2AGFP-W 8767 bp DNA circular SYN 15-DEC-2025
DEFINITION Cas9 activity reporter (control) with BFP and GFP..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8767)
AUTHORS Tzelepis K, Koike-Yusa H, De Braekeleer E, Li Y, Metzakopian E,
Dovey OM, Mupo A, Grinkevich V, Li M, Mazan M, Gozdecka M, Ohnishi
S, Cooper J, Patel M, McKerrell T, Chen B, Domingues AF, Gallipoli
P, Teichmann S, Ponstingl H, McDermott U, Saez-Rodriguez J, Huntly
BJ, Iorio F, Pina C, Vassiliou GS, Yusa K
TITLE A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and
Therapeutic Targets in Acute Myeloid Leukemia.
JOURNAL Cell Rep. 2016 Oct 18;17(4):1193-1205. doi:
10.1016/j.celrep.2016.09.079.
PUBMED 27760321
REFERENCE 2 (bases 1 to 8767)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 15, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.celrep.2016.09"; journalName: "Cell Rep"; date:
"2016-10-18- 18"; volume: "17"; issue: "4"; pages: "1193-1205"
COMMENT Sequence Label: pKLV2-U6gRNA5(Empty)-PGKBFP2AGFP-W
FEATURES Location/Qualifiers
source 1..8767
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(30..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(140..161)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 352..371
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 585..607
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 599..616
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 626..645
/label=T7
/note="T7 promoter, forward primer"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 960..1158
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1113..1133
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 1176..1356
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1403..1528
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 2027..2260
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2445..2489
/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 2787..2904
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2979..3227
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 2979..2999
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 3150..3169
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
promoter 3354..3853
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(3389..3412)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 3786..3805
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
regulatory 3877..3886
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 3883..4584
/codon_start=1
/product="monomeric blue fluorescent protein"
/label=TagBFP
/note="mammalian codon-optimized"
/translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
KLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
CDLPSKLGHKLN"
CDS 4603..4656
/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(4663..4665,4666..4668,4669..5382)
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS join(4663..4665,4666..4668,4669..5379)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4708..4729)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4969..4988)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 5316..5337
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(5442..5462)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5860..5871)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(6323..6340)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6329..6553
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(6579..6599)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(6579..6597)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(6618..6634)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6618..6634)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6631..6653)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6642..6658
/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(6666..6672,6673..6690,6691..6696))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6711..6732
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6849..6866)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7020..7608)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7100..7119)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(7779..8570,8571..8639))
/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 8402..8421
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8640..8744)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgactagtac
661 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
721 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
781 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
841 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
901 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
961 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
1021 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
1081 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
1141 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
1201 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
1261 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
1321 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
1381 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
1441 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
1501 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
1561 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
1621 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
1681 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
1741 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
1801 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
1861 gcaagcggcc ggccgcgctg atcttcagac ctggaggagg agatatgagg gacaattgga
1921 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
1981 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
2041 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
2101 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
2161 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
2221 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
2281 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
2341 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
2401 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
2461 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
2521 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
2581 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
2641 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
2701 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
2761 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
2821 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
2881 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
2941 ttagtaccgg gccctacgcg tccaaggtcg ggcaggaaga gggcctattt cccatgattc
3001 cttcatattt gcatatacga tacaaggctg ttagagagat aattagaatt aatttgactg
3061 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
3121 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
3181 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccggg tcttcgagaa
3241 gacacgttta agagctatgc tggaaacagc atagcaagtt taaataaggc tagtccgtta
3301 tcaacttgaa aaagtggcac cgagtcggtg ctttttttgg atccaattct accgggtagg
3361 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg
3421 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc
3481 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt
3541 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac
3601 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag
3661 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc
3721 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg
3781 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc
3841 tccgggcctt tcgacctgca tccatctaga tctcgagcca ccatgggcag cgagctgatt
3901 aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tggacaacca tcacttcaag
3961 tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg
4021 gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc
4081 agcaagacct tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct
4141 gagggcttca catgggagag agtcaccaca tacgaggacg ggggcgtgct gaccgctacc
4201 caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac
4261 ttcacatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgag
4321 accctgtacc ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg
4381 ggcgggagcc atctgatcgc aaacatcaag accacatata gatccaagaa acccgctaag
4441 aacctcaaga tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc
4501 aacaacgaga cctacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct
4561 agcaaactgg ggcacaagct taatggaggg ggcagcggcg gggaaggaag gggctccctg
4621 ctcacctgcg gcgacgtgga agagaacccc ggacctgaat tcatggtgag caagggcgag
4681 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
4741 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
4801 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
4861 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
4921 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
4981 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
5041 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
5101 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
5161 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
5221 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
5281 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
5341 gccgccggga tcactctcgg catggacgag ctgtacaagt aagcggccgc tcaacctctg
5401 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
5461 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
5521 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
5581 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
5641 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
5701 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
5761 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
5821 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
5881 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtctacg ccttcgccct
5941 cagacgagtc ggatctccct ttgggccgcc tccccgcagg tacctttaag accaatgact
6001 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
6061 attcactccc aaagaagtca agatctgctt tttgcctgta ctgggtctct ctggttagac
6121 cagagtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
6181 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
6241 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
6301 gcagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
6361 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
6421 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
6481 gtggggcagg acagcaaggg ggaggattgg gaagtcaata gcaggcatgc tggggatgcg
6541 gtgggctcta tgggcggccg ttaattaagc ttttgttccc tttagtgagg gttaattgcg
6601 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
6661 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
6721 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
6781 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
6841 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
6901 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
6961 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
7021 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
7081 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
7141 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
7201 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
7261 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
7321 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
7381 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
7441 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
7501 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
7561 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
7621 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
7681 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
7741 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
7801 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
7861 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
7921 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
7981 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
8041 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
8101 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
8161 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
8221 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
8281 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
8341 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
8401 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
8461 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
8521 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
8581 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
8641 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
8701 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
8761 gtgccac
//
