| 质粒名称: | pLentiCRISPR-RFP657 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 75162 |
| 存储实验室: | Beat Bornhauser |
|
载体骨架基本信息 |
|
| 载体名称: | lentiCRISPRv1 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 将嘌呤霉素抗性替换为RFP657 |
| 载体类型: | 哺乳动物表达，慢病毒，CRISPR |
| 筛选标记: | -- |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | RFP657 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pLentiCRISPR-RFP657 was a gift from Beat Bornhauser (Addgene plasmid # 75162 ; http://n2t.net/addgene:75162 ; RRID:Addgene_75162) |
| 参考文献部分: | Activation of concurrent apoptosis and necroptosis by SMAC mimetics for the treatment of refractory and relapsed ALL. McComb S, Aguade-Gorgorio J, Harder L, Marovca B, Cario G, Eckert C, Schrappe M, Stanulla M, von Stackelberg A, Bourquin JP, Bornhauser BC. Sci Transl Med. 2016 May 18;8(339):339ra70. doi: 10.1126/scitranslmed.aad2986. 10.1126/scitranslmed.aad2986 PubMed 27194728 |
买家导航
pLentiCRISPRRFP657载体基本信息
pLentiCRISPRRFP657载体图谱质粒图谱和多克隆位点信息
pLentiCRISPRRFP657载体简介
pLentiCRISPRRFP657载体序列
LOCUS pLentiCRISPR-RFP657 13540 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Derived from LentiCRISPR v1 but expresses RFP657 instead of
puromycin resistance.
ACCESSION .
VERSION .
KEYWORDS pLentiCRISPR-RFP657
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13540)
AUTHORS McComb S, Aguade-Gorgorio J, Harder L, Marovca B, Cario G, Eckert C,
Schrappe M, Stanulla M, von Stackelberg A, Bourquin JP, Bornhauser
BC
TITLE Activation of concurrent apoptosis and necroptosis by SMAC mimetics
for the treatment of refractory and relapsed ALL.
JOURNAL Sci Transl Med. 2016 May 18;8(339):339ra70. doi:
10.1126/scitranslmed.aad2986.
PUBMED 27194728
REFERENCE 2 (bases 1 to 13540)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..13540
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 5..231
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 232..412
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 459..584
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1077..1310
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1495..1539
/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"
promoter 1717..1957
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 1717..1737
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 1888..1907
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 3847..3922
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
misc_feature 3970..4087
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 4156..4367
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
regulatory 4387..4396
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4396..4419
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 4426..4446
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4471..8571
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 8572..8619
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 8629..8685
/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"
misc_feature 9400..9988
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(9453..9473)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(9871..9882)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 10057..10290
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 10368..10489
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(10405..10424)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 10459..10478
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 10529..10664
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 10591..10610
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(10684..10703)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(10685..10703)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(10713..10730)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(10713..10729)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10722..10744)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 10871..11326
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(10958..10977)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 11168..11189
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 11352..11456
/gene="bla"
/label=AmpR promoter
CDS 11457..12317
/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 complement(11675..11694)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 12488..13076
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 12977..12996
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 13230..13247
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 13364..13385
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 13400..13430
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 13438..13454
/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)."
primer_bind 13443..13465
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 13462..13478
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 13462..13478
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 13497..13517
/label=T3
/note="T3 promoter, forward primer"
promoter 13499..13517
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
1681 tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct
1741 tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta
1801 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
1861 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
1921 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggaga cggttgtaaa
1981 tgagcacaca aaatacacat gctaaaatat tatattctat gacctttata aaatcaacca
2041 aaatcttctt tttaataact ttagtatcaa taattagaat ttttatgttc ctttttgcaa
2101 acttttaata aaaatgagca aaataaaaaa acgctagttt tagtaactcg cgttgttttc
2161 ttcaccttta ataatagcta ctccaccact tgttcctaag cggtcagctc ctgcttcaat
2221 cattttttga gcatcttcaa atgttctaac tccaccagct gctttaacta aagcattgtc
2281 tttaacaact gacttcatta gtttaacatc ttcaaatgtt gcacctgatt ttgaaaatcc
2341 tgttgatgtt ttaacaaatt ctaatccagc ttcaacagct atttcacaag ctttcatgat
2401 ttcttctttt gttaataaac aattttccat aatacattta acaacatgtg atccagctgc
2461 tttttttaca gctttcatgt cttctaaaac taattcataa tttttgtctt ttaatgcacc
2521 aatatttaat accatatcaa tttctgttgc accatcttta attgcttcag aaacttcgaa
2581 tgcttttgta gctgttgtgc atgcacctag aggaaaacct acaacatttg ttattcctac
2641 atttgtgcct tttaataatt ctttacaata gcttgttcaa tatgaattaa cacaaactgt
2701 tgcaaaatca aattcaattg cttcatcaca taattgttta atttcagctt tcgtagcatc
2761 ttgttttaat aatgtgtgat ctatatattt gtttagtttc attttttctc ctatatattc
2821 atttttaatt ttaattcttt aataatttcg tctactttaa ctttagcgtt ttgaacagat
2881 tcaccaacac ctataaaata aatttttagt ttaggttcag ttccacttgg gcgaacagca
2941 aatcatgact tatcttctaa ataaaatttt agtaagtctt gtcctggcat attatacatt
3001 ccatcgatgt agtcttcaac attaacaact ttaagtccag caatttgagt taagggtgtt
3061 gctctcaatg atttcattaa tggttcaatt tttaatttct tttcttctgg tttaaaattc
3121 aagtttaaag tgaaagtgta atatgcaccc atttctttaa ataaatcttc taaatagtct
3181 actaatgttt tattttgttt tttataaaat caagcagcct ctgctattaa tatagaagct
3241 tgtattccat ctttatctct agctgagtca tcaattacat atccataact ttcttcataa
3301 gcaaaaacaa aatttaatcc gttatcttct tctttagcaa tttctctacc cattcattta
3361 aatccagtta aagtttttac aatattaact ccatattttt catgagcgat tctatcaccc
3421 aaatcacttg ttacaaaact tgaatataga gccggatttt ttggaatgct atttaagcgt
3481 tttagatttg ataattttca atcaattaaa attggtcctg tttgatttcc atctaatctt
3541 acaaaatgac catcatgttt tattgccatt ccaaatctgt cagcatctgg gtcattcata
3601 ataataatat ctgcatcatg tttaatacca tattcaagcg gtatttttca tgcaggatca
3661 aattctggat ttggatttac aacattttta aatgtttcat cttcaaatgc atgctcttca
3721 acctcaataa cgttatatcc tgattcacgt aatatttttg gggtaaattt agttcctgtt
3781 ccattaactg cgctaaaaat aatttttaaa tcttttttag cttcttgctc ttttttgtac
3841 gtctctgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
3901 aaaagtggca ccgagtcggt gcttttttga attctagatc ttgagacaaa tggcagtatt
3961 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
4021 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
4081 aaattttcgg gtttattaca gggacagcag agatccactt tggcgccggc tcgagtggct
4141 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
4201 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
4261 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
4321 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggtg tcgtgacgcg
4381 ggatccgcca ccatggatta caaagacgat gacgataaga tggccccaaa gaagaagcgg
4441 aaggtcggta tccacggagt cccagcagcc gacaagaagt acagcatcgg cctggacatc
4501 ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa
4561 ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg
4621 ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga
4681 tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc
4741 aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag
4801 aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag
4861 taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg
4921 cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag
4981 ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc
5041 tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc
5101 ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc
5161 gagaagaaga atggcctgtt cggcaacctg attgccctga gcctgggcct gacccccaac
5221 ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac
5281 gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg
5341 gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag
5401 atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac
5461 ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc
5521 ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag
5581 ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg
5641 aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc
5701 caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca
5761 ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac
5821 gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa
5881 accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc
5941 atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac
6001 agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc
6061 gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg
6121 ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa
6181 atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg
6241 ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa
6301 aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg
6361 atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg
6421 aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg
6481 gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga
6541 aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc
6601 caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc
6661 gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg
6721 ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag
6781 aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg
6841 ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg
6901 tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac
6961 cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc
7021 atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc
7081 tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg
7141 attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg
7201 gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg
7261 gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg
7321 gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag
7381 ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc
7441 gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc
7501 gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag
7561 gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc
7621 ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag
7681 atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa
7741 gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg
7801 cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac
7861 ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag
7921 ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga
7981 agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa
8041 aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag
8101 agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa
8161 tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat
8221 aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag
8281 cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg
8341 tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac
8401 ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc
8461 gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc
8521 atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga caagcgtcct
8581 gctgctacta agaaagctgg tcaagctaag aaaaagaaag ctagcggcgc aacaaacttc
8641 tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgagcga gctgattaag
8701 gagaacatgc acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc
8761 acatccgagg gcgaaggcaa gccctacgag ggcacccaga cccagagaat caaggtggtc
8821 gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagt
8881 cacaccttca tcaaccacac ccagggcatc cccgacttct ggaagcagtc cttccctgag
8941 ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag
9001 gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc
9061 ccatccaacg gccctgtgat gcagaagaaa acactcggct gggaggccca caccgagatg
9121 ctgtaccccg ctgacggcgg cctggaaggc agaaccgcgc tggccctgaa gctcgtgggc
9181 gggggccacc tgatctgcaa cttcaagacc acatacagat ccaagaaacc cgctaagaac
9241 ctcaagatgc ccggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccgac
9301 aaagagacct acgtcgagca gcacgaggtg gctgtggcca gatactgcga cctccctagc
9361 aaactggggc acaagcttaa ttaaacgcgt taagtcgaca atcaacctct ggattacaaa
9421 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
9481 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
9541 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
9601 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
9661 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
9721 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
9781 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
9841 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
9901 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
9961 cggatctccc tttgggccgc ctccccgcga ctttaagacc aatgacttac aaggcagctg
10021 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
10081 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
10141 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
10201 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
10261 ccttttagtc agtgtggaaa atctctagca gtacgtatag tagttcatgt catcttatta
10321 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
10381 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
10441 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
10501 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
10561 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
10621 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat
10681 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
10741 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
10801 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
10861 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
10921 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
10981 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
11041 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
11101 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
11161 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
11221 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
11281 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
11341 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
11401 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
11461 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
11521 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
11581 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
11641 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
11701 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
11761 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
11821 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
11881 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
11941 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
12001 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
12061 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
12121 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
12181 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
12241 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
12301 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
12361 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
12421 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
12481 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
12541 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
12601 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
12661 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
12721 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
12781 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
12841 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
12901 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
12961 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
13021 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
13081 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
13141 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
13201 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
13261 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
13321 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
13381 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
13441 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
13501 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
//
