| 质粒名称: | pSF4 TetCMV intron 20xGCN4 Renilla FKBP Stop 24xMS2v5 SV40 CTE polyA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 119945 |
| 存储实验室: | Jeffrey Chao |
|
载体骨架基本信息 |
|
| 载体名称: | pSF4 |
| 空载体大小: | 10500 bp |
| 完整质粒大小: | 11202 bp |
| 载体类型: | 哺乳动物表达，细菌表达 |
| 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: | Renilla luciferase |
| 物种来源: | Renilla reniformis |
| 插入大小: | 930 bp |
| 启动子: | TetCMV |
| 标签/融合蛋白: |
24xGCN4重复（SunTag）（插入件上的N端子）
FKBP（插入件上的C端子）
24xMS2v5 stem loops in 3'UTR
|
| 5’测序引物 | AATACGACTCACTATAGGC |
|
文献引用方法 |
|
| 材料和方法部分: | pSF4 TetCMV intron 20xGCN4 Renilla FKBP Stop 24xMS2v5 SV40 CTE polyA was a gift from Jeffrey Chao (Addgene plasmid # 119945 ; http://n2t.net/addgene:119945 ; RRID:Addgene_119945) |
| 参考文献部分: | Single-Molecule Imaging of mRNA Localization and Regulation during the Integrated Stress Response. Wilbertz JH, Voigt F, Horvathova I, Roth G, Zhan Y, Chao JA. Mol Cell. 2019 Jan 4. pii: S1097-2765(18)31036-0. doi: 10.1016/j.molcel.2018.12.006. 10.1016/j.molcel.2018.12.006 PubMed 30661979 |
pSF4 TetCMV intron 20xGCN4 Renilla FKBP Stop 24xMS2v5 SV40 C
买家导航
pSF4 TetCMV intron 20xGCN4 Renilla FKBP Stop 24xMS2v5 SV40 C载体质粒基本信息
pSF4 TetCMV intron 20xGCN4 Renilla FKBP Stop 24xMS2v5 SV40 C质粒图谱载体图谱和pSF4 TetCMV intron 20xGCN4 Renilla FKBP Stop 24xMS2v5 SV40 C载体序列质粒序列多克隆位点信息
pSF4 TetCMV intron 20xGCN4 Renilla FKBP Stop 24xMS2v5 SV40 C质粒载体简介
pSF4 TetCMV intron 20xGCN4 Renilla FKBP Stop 24xMS2v5 SV40 C质粒序列载体序列
LOCUS pSF4 TetCMV intron 20xGCN4 Renilla FKBP Stop 24xMS2v5 SV40 CTE... 11192 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses SunTag-Renilla mRNA reporter in mammalian cells; note
plasmid contains 24xGCN4 repeats.
ACCESSION .
VERSION .
KEYWORDS pSF4 TetCMV intron 20xGCN4 Renilla FKBP Stop 24xMS2v5 SV40 CTE...
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11192)
AUTHORS Wilbertz JH, Voigt F, Horvathova I, Roth G, Zhan Y, Chao JA
TITLE Single-Molecule Imaging of mRNA Localization and Regulation during
the Integrated Stress Response.
JOURNAL Mol Cell. 2019 Jan 4. pii: S1097-2765(18)31036-0. doi:
10.1016/j.molcel.2018.12.006.
PUBMED 30661979
REFERENCE 2 (bases 1 to 11192)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..11192
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(12..28)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(12..28)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(25..47)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 36..52
/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(60..90)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 105..126
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(243..260)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(414..1002)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(494..513)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1173..2033)
/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 1796..1815
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2034..2138)
/gene="bla"
/label=AmpR promoter
primer_bind 2206..2224
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2262..2284)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2384..2403
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2597..2619
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2611..2628
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2612..2628
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3710..3757)
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
protein_bind 3804..3822
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3840..3858
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3876..3894
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3912..3930
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3948..3966
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3984..4002
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4020..4038
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 4083..4107
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 4184..4316
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 4299..4318
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter 4361..4379
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 4383..4392
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4458..4514
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 4530..4586
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 4602..4658
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 4674..4730
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 4746..4802
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 4818..4874
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 4890..4946
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 4962..5018
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5034..5090
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5106..5162
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5178..5234
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5250..5306
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5322..5378
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5394..5450
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5466..5522
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5538..5594
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5610..5666
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5682..5738
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5754..5810
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5826..5882
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5898..5954
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 5970..6026
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 6114..6170
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 6204..7133
/codon_start=1
/product="Renilla luciferase"
/label=hRluc
/note="human codon-optimized"
/translation="ASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENA
VIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWF
ELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIA
LIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREI
PLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVK
GLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
CDS 7134..7457
/codon_start=1
/product="destabilization domain that can be stabilized by
Shield1 in the ProteoTuner(TM) system"
/label=DD
/note="L106P mutant of FKBP12"
/translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
LKPE"
polyA_signal 8990..9124
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind 9068..9087
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(9318..9334)
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
primer_bind complement(9350..9372)
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
misc_feature 9783..10140
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
LTR 10205..10674
/note="long terminal repeat from Moloney murine leukemia
virus"
ORIGIN
1 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
61 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
121 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
181 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
241 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
301 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
361 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
421 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
481 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
541 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
601 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
661 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
721 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
781 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
841 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
901 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
961 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
1021 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt tgtcatgaga
1081 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
1141 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
1201 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
1261 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
1321 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
1381 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
1441 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
1501 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
1561 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
1621 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
1681 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
1741 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
1801 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
1861 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
1921 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
1981 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
2041 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
2101 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
2161 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
2221 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
2281 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
2341 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
2401 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
2461 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
2521 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
2581 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attaattcca
2641 agctaagtga aagactctgt ctcaaaaacc aacctgacac ctgagccgga gcgttgcctc
2701 tcactaccat aaatatatat acacaacaca cacacacaca cacacacaca cacacacaca
2761 cacacacaca atcatacaca taaacacaat cacacacaca cacatgcaca atcacacaca
2821 cacaatcaca cacaaacata catacacaca atcacacaca accatacaca catatacaca
2881 atcacacaca caaacacaat cacacacaca cacatcaatc aatcaatcaa tcaatatacc
2941 aaaaatgata gctcatacct gtaatcttag atgtgggaag gctaaaacag caggctaccc
3001 atgacttcaa ggtcaccctg ggttacagag tagctaatag tctagcttga actatagtgt
3061 aggatcttta aaaaaaaatt aaccaaccaa ccaacaaaca aacaaacaaa caacaacaac
3121 aacaaatgga agacagagga caaaaaaaaa taaaagaaaa agtagaaagc caatggagat
3181 actatcttaa aattagaaat atttatgaca cattctgaat actccatgtt tatttcaagg
3241 aagccaatgt aactttaagt actatggcca tttaatgtaa atacttaaga aaaaaaacca
3301 aattaatttt gatacatgct gcatgtgaaa gacccccgct gacgggtagt caatcactca
3361 gaggagaccc tctcaaggaa cagcgagacc acaagtcgga tgcaactgca agagggttta
3421 ttggatacac gggtacccgg gcgactcagt caatcggagg actggcgccc cgagtgaggg
3481 gttgtgggct cgagctcggg gagcagaagc gcgcgaacag aagtcgagct agcttgccaa
3541 acctacaggt ggggtctttc attcccccct ttttctggag actaaataaa atcttttatt
3601 ttatctatgg ctcgtactct ataggcttca gctggtgata ttgttgagtc aaaactagag
3661 cctggaccac tgatatcctg tctttaacaa attggactaa tccggatctg aagttcctat
3721 actttctaga gaataggaac ttcggaatag gaacttcgcg gccagatctt ttgcggccgc
3781 tttgggccgg ccgaattctt tactccctat cagtgataga gaacgtatga agagtttact
3841 ccctatcagt gatagagaac gtatgcagac tttactccct atcagtgata gagaacgtat
3901 aaggagttta ctccctatca gtgatagaga acgtatgacc agtttactcc ctatcagtga
3961 tagagaacgt atctacagtt tactccctat cagtgataga gaacgtatat ccagtttact
4021 ccctatcagt gatagagaac gtataagctt taggcgtgta cggtgggcgc ctataaaagc
4081 agagctcgtt tagtgaaccg tcagatcgcc tggagcaatt ccacaacact tttgtcttat
4141 accaactttc cgtaccactt cctaccctcg taaaaccggt caggtaagta tcaaggttac
4201 aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg
4261 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt
4321 ccactcccag ttcaattaca gctcttaagg ctagagtact taatacgact cactataggc
4381 tagccaccat gggcgccatc atacacgcgg ccgcaatcat cggtccgggt ggatctggag
4441 gtggaggttc tggaggagaa gaacttttga gcaagaatta tcatcttgag aacgaagtgg
4501 ctcgtcttaa gaaaggttct ggcagtggag aagaactgct ttcaaagaat taccacctgg
4561 aaaatgaggt agctagactg aaaaagggga gcggaagtgg ggaggagttg ctgagcaaaa
4621 attatcattt ggagaacgaa gtagcacgac taaagaaagg gtccggatcg ggtgaggagt
4681 tactctcgaa aaattatcat ctcgaaaacg aagtggctcg gctaaaaaag ggcagtggtt
4741 ctggagaaga gctattatct aaaaactacc acctcgaaaa tgaggtggca cgcttaaaaa
4801 agggaagtgg cagtggtgaa gagctactat ccaagaatta tcatcttgag aacgaggtag
4861 cgcgtttgaa gaagggttcc ggctcaggag aggaactgct ctcgaagaac tatcatcttg
4921 aaaatgaggt cgctcgatta aaaaagggat cgggcagtgg tgaggaacta ctttcaaaga
4981 attaccacct cgaaaacgaa gtagctcgat taaagaaagg ttcagggtcg ggtgaagaat
5041 tactgagtaa aaattatcat ctggaaaatg aggtagcgag actaaaaaag gggagtggtt
5101 ctggcgaaga gttgctatcg aaaaattatc atcttgagaa cgaagttgct aggctcaaaa
5161 agggctcagg ctcaggcgag gagttgctct cgaaaaacta ccacttggaa aatgaggtcg
5221 cgaggttgaa aaaggggagc gggtcgggcg aggagttatt gagcaaaaac tatcatttag
5281 agaacgaagt cgcgcgctta aagaaaggct cgggctcggg cgaagaactc ttatcgaaga
5341 actaccacct cgaaaatgag gtcgccaggt tgaaaaaggg cagtggcagc ggggaggaac
5401 tcttgagcaa gaactaccac ttggagaatg aggtcgcgag attgaagaaa gggtcgggga
5461 gcggcgagga attgctcagc aagaattatc atttggagaa cgaagtcgcc aggctcaaga
5521 aaggctcggg gtcgggggag gaattgttga gtaaaaacta ccacttggaa aatgaagtcg
5581 ccaggctcaa aaaagggagt gggagcggcg aagagttatt gagcaaaaat taccacttgg
5641 agaacgaagt ggcaaggctc aagaaaggga gcggcagcgg ggaggagctc ttatcgaaga
5701 actaccactt agagaatgaa gtcgcccgct tgaagaaagg ctcggggagc ggggaagagc
5761 tcttgagcaa gaactaccac ttggaaaatg aggtggcgcg cttgaagaaa gggagcggga
5821 gcggggaaga gttactatct aagaattatc atctcgagaa cgaggtggct cgactaaaga
5881 agggctccgg cagtggggag gaactcctgt cgaagaacta tcatcttgaa aatgaggttg
5941 caagacttaa aaaggggtcc ggatcaggtg aggaactact cagtaagaat taccacctgg
6001 aaaacgaagt agctcgatta aagaaaggtt cagggtcggg tgaagaactg ctctcaaaag
6061 attatcattt ggaaaatgag gttgcacgtt taaaaaaggg aagtggcagt ggtgaggaac
6121 ttctgtcgaa aaattatcat ctcgagaatg aagtagcccg acttaaaaag ggttctggct
6181 cgggtcagcg gccgcaccgg tcagcttcca aggtgtacga ccccgagcaa cgcaaacgca
6241 tgatcactgg gcctcagtgg tgggctcgct gcaagcaaat gaacgtgctg gactccttca
6301 tcaactacta tgattccgag aagcacgccg agaacgccgt gatttttctg catggtaacg
6361 ctgcctccag ctacctgtgg aggcacgtcg tgcctcacat cgagcccgtg gctagatgca
6421 tcatccctga tctgatcgga atgggtaagt ccggcaagag cgggaatggc tcatatcgcc
6481 tcctggatca ctacaagtac ctcaccgctt ggttcgagct gctgaacctt ccaaagaaaa
6541 tcatctttgt gggccacgac tggggggctt gtctggcctt tcactactcc tacgagcacc
6601 aagacaagat caaggccatc gtccatgctg agagtgtcgt ggacgtgatc gagtcctggg
6661 acgagtggcc tgacatcgag gaggatatcg ccctgatcaa gagcgaagag ggcgagaaaa
6721 tggtgcttga gaataacttc ttcgtcgaga ccatgctccc aagcaagatc atgcggaaac
6781 tggagcctga ggagttcgct gcctacctgg agccattcaa ggagaagggc gaggttagac
6841 ggcctaccct ctcctggcct cgcgagatcc ctctcgttaa gggaggcaag cccgacgtcg
6901 tccagattgt ccgcaactac aacgcctacc ttcgggccag cgacgatctg cctaagatgt
6961 tcatcgagtc cgaccctggg ttcttttcca acgctattgt cgagggagct aagaagttcc
7021 ctaacaccga gttcgtgaag gtgaagggcc tccacttcag ccaggaggac gctccagatg
7081 aaatgggtaa gtacatcaag agcttcgtgg agcgcgtgct gaagaacgag cagatgggag
7141 tgcaggtgga aaccatctcc ccaggagacg ggcgcacctt ccccaagcgc ggccagacct
7201 gtgtggtgca ctacaccggg atgcttgaag atggaaagaa agtcgattcc tcccgggaca
7261 gaaacaagcc ctttaagttt atgctaggca agcaggaggt gatccgaggc tgggaagaag
7321 gggttgccca gatgagtgtg ggtcagagag ccaaactgac tatatctcca gattatgcct
7381 atggtgccac tgggcaccca ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg
7441 agcttctaaa accggaataa gggccaggga agaactggct cggaaagagg accccacatg
7501 ttgtagcttc aagtttaaac ggatccaacc tacaaacggg tggaggatca ccccacccga
7561 cacttcacaa tcaaggggta caatacacaa gggtggagga acaccccacc ctccagacac
7621 attacacaga aatccaatca aacagaagca ccatcagggc ttctgctacc aaatttatct
7681 caaaaaacta caacaaggaa tcaccatcag ggattccctg tgcaatatac gtcaaacgag
7741 ggccacgacg ggaggacgat cacgcctccc gaatatcggc atgtctggct ttcgaattca
7801 gtgcgtggag catcagccca cgcagccaat cagagtcgaa tacaagtcga ctttcgcgaa
7861 gagcatcagc cttcgcgcca ttcttacaca aaccacactc tcccctacag gaacagcatc
7921 agcgttcctg cccagtaccc aactcaagaa aatttatgtc cccatgcagc atcagcgcat
7981 gggccccaag aatacatccc caacaaaatc acatccgagc accaacaggg ctcggagtgt
8041 tgtttcttgt ccaactggac aaaccctcca tggaccatca ggccatggac tctcaccaac
8101 aagacaaaaa ctactcttct cgaagcagca tcagcgcttc gaaacactcg agcatacatt
8161 gtgcctattt cttgggtgga cgatcacgcc acccatgctc tcacgaattt caaaacacgg
8221 acaaggacga gcaccaccag ggctcgtcgt tccacgtcca atacgattac ttacctttcg
8281 ggatcacgat cacggatccc gcagctacat cacttccact caggacattc aagcatgcac
8341 gatcacggca tgctccacaa gtctcaacca cagaaactac caaatgggtt cagcaccagc
8401 gaacccactc ctacctcaaa cctcttccca caaaactggc aagcaggatc accgcttgcc
8461 cattccaaca taccaaatca aaaacaatta ctggtacagc atcagcgtac cagcccacat
8521 ctctcactac tatcaaaaac caaaccgttc agcaacagcg aacggtacac acggaaaaat
8581 caactggttt acaaatacga aagacgatca cgctttcgtc cagcgcaaac tattacgaaa
8641 aacatccgac gggaagagca acagccttcc cgcggcggaa aacctcacaa aaacacgaca
8701 aacggatgca cgaacacggc atccgccgac aacccacaaa cttacaacca ggcaaacggt
8761 gcaggatcac cgcaccgtac atcaaacacc tcagatctca tttcgcgcgg gatccagacc
8821 acctcccctg cgagctaagc tggacagcca atgacgggta agagagtgac atttttcact
8881 aacctaagac aggagggccg tcagagctac tgcctaatcc aaagacgggt aaaagtgata
8941 aaaatgtatc actccaacct aagacaggcg cagcttccga gggatttgag atccagacat
9001 gataagatac attgatgagt ttggacaaac caaaactaga atgcagtgaa aaaaatgcct
9061 tatttgtgaa atttgtgatg ctattgcctt atttgtaacc attataagct gcaataaaca
9121 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
9181 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc tgaagttcct
9241 atactatttg aagaatagga acttcggaat aggaacttcg gatctcatat ggccatatgg
9301 gggcgcctag agaaggagtg agggctggat aaagggagga tcgaggcggg gtcgaacgag
9361 gaggttcaag ggggagagac ggggcggatg gaggaagagg aggcggaggc ttagggtgta
9421 caaagggctt gacccaggga ggggggtcaa aagccaaggc ttcccaggtc acgatgtagg
9481 ggacctggtc tgggtgtcca tgcgggccag gtgaaaagac cttgatctta acctgggtga
9541 tgaggtctcg gttaaaggtg ccgtctcgcg gccatccgac gttaaaggtt ggccattctg
9601 cagagcagaa ggtaacccaa cgtctcttct tgacatctac cgactggttg tgagcgatcc
9661 gctcgacatc tttccagtga cctaaggtca aacttaaggg agtggtaaca ggctagcccg
9721 ggcccatatt ttcagacaaa tacagaaaca cagtcagaca gagacaacac agaacgatgc
9781 tgcagcagac aagacgcgcg gcgcggcttc ggtcccaaac cgaaagcaaa aattcagacg
9841 gaggcgggaa ctgttttagg ttctcgtctc ctaccagaac cacatatccc tcctctaagg
9901 ggggtgcacc aaagagtcct aaacgatcgg gattttagga ctcaggtcgg gccacaaaaa
9961 cggcccccga agtccctggg acgtctccca gggttgcggc cgggtgttcc gaactcgtca
10021 gttccaccac gggtccgcca gatacagagc tagttagcta actagtaccg acgcaggcgc
10081 ataaaatcag tcatagacac tagacaatcg gacagacaca gataagttgc tggccagctt
10141 acctcccggt ggtgggtcgg tggtccctgg gcaggggtct cccgatcccg gacgagcccc
10201 caaatgaaag acccccgctg acgggtagtc aatcactcag aggagaccct cccaaggaac
10261 agcgagacca caagtcggat gcaactgcaa gagggtttat tggatacacg ggtacccggg
10321 cgactcagtc aatcggagga ctggcgcgcc gagtgagggg ttgtgggctc ttttattgag
10381 ctcggggagc agaagcgcgc gaacagaagc gagaagcgaa ctgattggtt agttcaaata
10441 aggcacaggg tcatttcagg tccttggggc accctggaaa catctgatgg ttctctagaa
10501 actgctgagg gctggaccgc atctggggac catctgttct tggccctgag ccggggcagg
10561 aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct gttcttggcc
10621 ctgagccggg gcaggaactg cttaccacag atatcctgtt tggcccatat tcagctgttc
10681 catctgttcc tgaccttgat ctgaactttt ctattctcag ttatgtattt ttccatgcct
10741 tgcaaaatgg cgttacttaa gctagcttgc caaacctaca ggtggggtct ttcacatgta
10801 tatgtcaaaa ataaaaatca actaattgac tagtaattaa tatgactggc ataatgggaa
10861 attgatcctg acagatgcaa actggcttct cagcagcgca tttatgttgt caactgagga
10921 aggaaacgtt aatgacagaa actctaagta atttccacgt ttatctattt ttatttatac
10981 tagctttggt aacaggaata ttgcagcatt catgcacatt gaaaccctta tgaaataaaa
11041 acatctgtgc atttaaaatg gaattaacat tttaaatgtt aaaaaaagct ggcttagctt
11101 ccccccgccc cctagggcat agaacaagtc aaatgcttta tatatttgag tttgggatgt
11161 attaggaaac tcctaagagc aaagctagct tg
//
