| 质粒名称: | pLenti-HF1RA-P2A-GFP-PGK-Puro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 110866 |
| 存储实验室: | Lukas Dow |
|
载体骨架基本信息 |
|
| 载体名称: | Adapted from lentiCRISPRv1 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6296 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 慢病毒表达 |
| 筛选标记: | 嘌呤霉素；GFP |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Cas9-HF1RA,HF1RA |
| 物种来源: | Synthetic |
| 插入大小: | 4227 bp |
| 基因突变: | R661A, Q695A, Q926A, and NLS sequence at the N-terminus |
| GenBank编号 | -- |
| 启动子 | EF1s |
| 标签/融合蛋白: | FLAG（插入N端子） |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | TAAGTGCAGTAGTCGCCGTG |
| 3' 测序引物 | TTAAGAATACCAGTCAATCTTTC |
|
文献引用方法 |
|
| 材料和方法部分: | pLenti-HF1RA-P2A-GFP-PGK-Puro was a gift from Lukas Dow (Addgene plasmid # 110866 ; http://n2t.net/addgene:110866 ; RRID:Addgene_110866) |
| 参考文献部分: | Optimized base editors enable efficient editing in cells, organoids and mice. Zafra MP, Schatoff EM, Katti A, Foronda M, Breinig M, Schweitzer AY, Simon A, Han T, Goswami S, Montgomery E, Thibado J, Kastenhuber ER, Sanchez-Rivera FJ, Shi J, Vakoc CR, Lowe SW, Tschaharganeh DF, Dow LE. Nat Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194. 10.1038/nbt.4194 PubMed 29969439 |
买家导航
pLenti-HF1RA-P2A-GFP-PGK-Puro载体质粒基本信息
pLenti-HF1RA-P2A-GFP-PGK-Puro质粒图谱载体图谱和pLenti-HF1RA-P2A-GFP-PGK-Puro载体序列质粒序列多克隆位点信息
pLenti-HF1RA-P2A-GFP-PGK-Puro质粒载体简介
pLenti-HF1RA-P2A-GFP-PGK-Puro质粒序列载体序列
LOCUS pLenti-HF1RA-P2A-GFP-PGK-Puro 12448 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Lentiviral vector for constitutive expression of Cas9-HF1RA-P2A-GFP
in mammalian cells (codon optimized).
ACCESSION .
VERSION .
KEYWORDS pLenti-HF1RA-P2A-GFP-PGK-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12448)
AUTHORS Zafra MP, Schatoff EM, Katti A, Foronda M, Breinig M, Schweitzer AY,
Simon A, Han T, Goswami S, Montgomery E, Thibado J, Kastenhuber ER,
Sanchez-Rivera FJ, Shi J, Vakoc CR, Lowe SW, Tschaharganeh DF, Dow
LE
TITLE Optimized base editors enable efficient editing in cells, organoids
and mice.
JOURNAL Nat Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194.
PUBMED 29969439
REFERENCE 2 (bases 1 to 12448)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..12448
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 222..402
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 449..574
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1067..1300
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 1713..1830
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1899..2110
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
regulatory 2130..2139
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2139..2162
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2169..2189
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2214..6314
/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
KTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFMA
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 6315..6362
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 6372..6428
/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 6429..7148
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(6474..6495)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(6735..6754)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 7082..7103
/label=EGFP-C
/note="EGFP, forward primer"
promoter 7160..7659
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(7195..7218)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 7592..7611
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 7680..8279
/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(7680..7699)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 8176..8196
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 8295..8883
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(8348..8368)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(8766..8777)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 8955..9188
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 9266..9387
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9303..9322)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 9357..9376
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 9427..9562
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 9489..9508
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(9582..9601)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(9583..9601)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(9611..9628)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(9611..9627)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9620..9642)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 9769..10224
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(9856..9875)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 10066..10087
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 10250..10354
/gene="bla"
/label=AmpR promoter
CDS 10355..11215
/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(10573..10592)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 11386..11974
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 11875..11894
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 12128..12145
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 12262..12283
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 12298..12328
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 12336..12352
/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 12341..12363
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 12360..12376
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 12360..12376
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 12395..12415
/label=T3
/note="T3 promoter, forward primer"
promoter 12397..12415
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 12443..221
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
ORIGIN
1 cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac
61 aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct
121 tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt
181 gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc
241 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
301 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
361 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
421 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
481 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
541 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
601 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
661 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
721 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
781 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
841 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
901 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
961 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
1021 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
1081 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
1141 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
1201 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
1261 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
1321 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
1381 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
1441 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
1501 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
1561 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
1621 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
1681 acctcccaac cccgagggga cccagaccac aattttaaaa gaaaaggggg gattgggggg
1741 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
1801 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
1861 ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
1921 agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg
1981 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
2041 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
2101 cagaacacag gtgtcgtgac gcgggatccg ccaccatgga ttacaaagac gatgacgata
2161 agatggcccc aaagaagaag cggaaggtcg gtatccacgg agtcccagca gccgacaaga
2221 agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg
2281 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
2341 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
2401 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
2461 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
2521 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
2581 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
2641 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
2701 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
2761 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
2821 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
2881 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc
2941 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
3001 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
3061 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
3121 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
3181 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
3241 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg
3301 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
3361 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
3421 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
3481 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
3541 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
3601 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
3661 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
3721 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc
3781 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
3841 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
3901 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
3961 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
4021 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
4081 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
4141 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc gccctgagcc
4201 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
4261 agtccgacgg cttcgccaac agaaacttca tggccctgat ccacgacgac agcctgacct
4321 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
4381 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
4441 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
4501 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
4561 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
4621 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
4681 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc
4741 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc
4801 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
4861 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg
4921 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
4981 cccgggccat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
5041 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
5101 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
5161 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
5221 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
5281 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
5341 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
5401 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc
5461 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
5521 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga
5581 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
5641 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
5701 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
5761 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
5821 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
5881 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
5941 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc
6001 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
6061 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag
6121 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
6181 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
6241 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
6301 agctgggagg cgacaagcgt cctgctgcta ctaagaaagc tggtcaagct aagaaaaaga
6361 aaggcagcgg cgccaccaac ttcagcctgc tgaagcaggc cggcgacgtg gaggagaacc
6421 ccggccccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
6481 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg
6541 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
6601 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
6661 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
6721 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
6781 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
6841 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
6901 agaagaacgg catcaaggcg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
6961 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
7021 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
7081 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
7141 acaagtaagc tagcctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc
7201 gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat
7261 tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct
7321 tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg
7381 tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa
7441 tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg
7501 gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg
7561 gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct
7621 gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttacca
7681 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
7741 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
7801 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
7861 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
7921 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
7981 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
8041 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
8101 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
8161 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
8221 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgat
8281 tctagaaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
8341 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
8401 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
8461 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
8521 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
8581 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
8641 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
8701 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
8761 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
8821 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
8881 cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
8941 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
9001 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
9061 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
9121 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
9181 ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
9241 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
9301 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
9361 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
9421 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
9481 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
9541 tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt
9601 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
9661 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
9721 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
9781 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
9841 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
9901 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
9961 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
10021 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
10081 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
10141 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
10201 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
10261 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
10321 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
10381 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
10441 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
10501 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
10561 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
10621 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
10681 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
10741 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
10801 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
10861 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
10921 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
10981 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
11041 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
11101 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
11161 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
11221 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
11281 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
11341 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
11401 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
11461 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
11521 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
11581 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
11641 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
11701 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
11761 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
11821 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
11881 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
11941 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
12001 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
12061 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
12121 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
12181 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
12241 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
12301 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
12361 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
12421 aaagctggag ctgcaagctt aatgtagt
//
