| 质粒名称: | pSLIK-IDH1-R132H-FLAG |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 66803 |
| 存储实验室: | Christian Metallo |
|
载体骨架基本信息 |
|
| 载体名称: | pSLIK-hygro |
| 载体出品公司: | Ian Frasier, Addgene Plasmid #25737 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 13918 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒 |
| 筛选标记: | 潮霉素 |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | IDH1,isocitrate dehydrogenase |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1357 bp |
| 基因突变: | R132H |
| GenBank编号 | NM_005896.3 |
| 启动子 | CMV |
| 标签/融合蛋白: | 3xFLAG（插接件上的C端子） |
| 克隆方法: | Gateway克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | huB-Cpro-R |
|
文献引用方法 |
|
| 材料和方法部分: | pSLIK-IDH1-R132H-FLAG was a gift from Christian Metallo (Addgene plasmid # 66803 ; http://n2t.net/addgene:66803 ; RRID:Addgene_66803) |
| 参考文献部分: | Tracing compartmentalized NADPH metabolism in the cytosol and mitochondria of mammalian cells. Lewis CA, Parker SJ, Fiske BP, McCloskey D, Gui DY, Green CR, Vokes NI, Feist AM, Vander Heiden MG, Metallo CM. Mol Cell. 2014 Jul 17;55(2):253-63. doi: 10.1016/j.molcel.2014.05.008. Epub 2014 May 29. 10.1016/j.molcel.2014.05.008 PubMed 24882210 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pSLIK-IDH1-R132H-FLAG 13969 bp ds-DNA circular SYN 17-JAN-2019
DEFINITION Tet-inducible expression of mutant IDH1 in mammalian cells, 3rd
generation lentiviral vector.
ACCESSION .
VERSION .
KEYWORDS pSLIK-IDH1-R132H-FLAG
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13969)
AUTHORS Lewis CA, Parker SJ, Fiske BP, McCloskey D, Gui DY, Green CR, Vokes
NI, Feist AM, Vander Heiden MG, Metallo CM
TITLE Tracing compartmentalized NADPH metabolism in the cytosol and
mitochondria of mammalian cells.
JOURNAL Mol Cell. 2014 Jul 17;55(2):253-63. doi:
10.1016/j.molcel.2014.05.008. Epub 2014 May 29.
PUBMED 24882210
REFERENCE 2 (bases 1 to 13969)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 17, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..13969
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(11..28)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(182..770)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(262..281)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(941..1801)
/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 1564..1583
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1802..1906)
/gene="bla"
/label=AmpR promoter
primer_bind complement(1981..2000)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 2172..2551
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2553..2751
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2706..2726
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 2769..2949
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 2996..3121
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 3614..3847
/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 4374..4491
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 4548..4572
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
promoter 4604..4918
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
protein_bind 4612..4630
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4648..4666
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4683..4701
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4719..4737
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4755..4773
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4790..4808
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4826..4844
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 4891..4915
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 6194..6259
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
protein_bind complement(6280..6304)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
promoter 6354..7565
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 7402..7421
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
CDS 7690..8436
/codon_start=1
/product="improved tetracycline-controlled transactivator"
/label=rtTA-Advanced
/note="In the Tet-On(R) system, rtTA-Advanced binds to the
Tet-responsive element and stimulates transcription only in
the presence of tetracycline or doxycycline."
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
misc_feature 8474..9047
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(8641..8658)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 8868..8887
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 9106..10125
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
misc_feature 10591..11179
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(10644..10664)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(11062..11073)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(11182..11198)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(11183..11199)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 11708..11888
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(11914..11931)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 11920..12144
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 12190..12618
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(12277..12296)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 12487..12508
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(12627..12647)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 12632..12961
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 12812..12947
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 12874..12893
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 13009..13056
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 13075..13449
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 13579..13700
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(13616..13635)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 13670..13689
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(13749..13765)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(13749..13765)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(13762..13784)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 13773..13789
/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(13797..13827)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 13842..13863
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg
2761 cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
2821 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
2881 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
2941 tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc
3001 tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt
3061 gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt
3121 cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg
3181 aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc
3241 agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca
3301 accatccctt cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct
3361 ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag acaagataga
3421 ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg
3481 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
3541 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
3601 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
3661 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
3721 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
3781 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
3841 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
3901 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
3961 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
4021 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
4081 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
4141 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
4201 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg
4261 aaggaataga agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg
4321 gatcggcact gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa
4381 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
4441 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
4501 tacagggaca gcagagatcc agtttggtta attctagaca gttatcaaca agtttgtaca
4561 aaaaagcagg ctttaaagga accaattcag tcgactggat cccgagttta ctccctatca
4621 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacga tgtcgagttt
4681 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
4741 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttat ccctatcagt
4801 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgaggtag
4861 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgccac
4921 tagtgccacc atgtccaaaa aaatcagtgg cggttctgtg gtagagatgc aaggagatga
4981 aatgacacga atcatttggg aattgattaa agagaaactc atttttccct acgtggaatt
5041 ggatctacat agctatgatt taggcataga gaatcgtgat gccaccaacg accaagtcac
5101 caaggatgct gcagaagcta taaagaagca taatgttggc gtcaaatgtg ccactatcac
5161 tcctgatgag aagagggttg aggagttcaa gttgaaacaa atgtggaaat caccaaatgg
5221 caccatacga aatattctgg gtggcacggt cttcagagaa gccattatct gcaaaaatat
5281 cccccggctt gtgagtggat gggtaaaacc tatcatcata ggtcatcatg cttatgggga
5341 tcaatacaga gcaactgatt ttgttgttcc tgggcctgga aaagtagaga taacctacac
5401 accaagtgac ggaacccaaa aggtgacata cctggtacat aactttgaag aaggtggtgg
5461 tgttgccatg gggatgtata atcaagataa gtcaattgaa gattttgcac acagttcctt
5521 ccaaatggct ctgtctaagg gttggccttt gtatctgagc accaaaaaca ctattctgaa
5581 gaaatatgat gggcgtttta aagacatctt tcaggagata tatgacaagc agtacaagtc
5641 ccagtttgaa gctcaaaaga tctggtatga gcataggctc atcgacgaca tggtggccca
5701 agctatgaaa tcagagggag gcttcatctg ggcctgtaaa aactatgatg gtgacgtgca
5761 gtcggactct gtggcccaag ggtatggctc tctcggcatg atgaccagcg tgctggtttg
5821 tccagatggc aagacagtag aagcagaggc tgcccacggg actgtaaccc gtcactaccg
5881 catgtaccag aaaggacagg agacgtccac caatcccatt gcttccattt ttgcctggac
5941 cagagggtta gcccacagag caaagcttga taacaataaa gagcttgcct tctttgcaaa
6001 tgctttggaa gaagtctcta ttgagacaat tgaggctggc ttcatgacca aggacttggc
6061 tgcttgcatt aaaggtttac ccaatgtgca acgttctgac tacttgaata catttgagtt
6121 catggataaa cttggagaaa acttgaagat caaactagct caggccaaac tttattctag
6181 aggatcccgg gctgactaca aagaccatga cggtgattat aaagatcatg acatcgacta
6241 caaggatgac gatgacaagt agtgactcga gatatctaga cccagctttc ttgtacaaag
6301 tggttgataa catctcgagc cataaagatg gttaattaac ccacccaaga tctggcctcc
6361 gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcctc acggcgagcg ctgccacgtc
6421 agacgaaggg cgcagcgagc gtcctgatcc ttccgcccgg acgctcagga cagcggcccg
6481 ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt taggacggga
6541 cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg aggaaaagta
6601 gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc gatgattata
6661 taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt
6721 cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg gctggccggg
6781 gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac cgccaagggc
6841 tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga gcgcagcaaa
6901 atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt
6961 gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgaggc cttcgctaat
7021 gcgggaaagc tcttattcgg gtgagatggg ctggggcacc atctggggac cctgacgtga
7081 agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg
7141 gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt
7201 cacccgttct gttggcttat aatgcagggt ggggccacct gccggtaggt gtgcggtagg
7261 cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg
7321 cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat
7381 gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga
7441 gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta
7501 attttcagtg ttagactagt aaattgtccg ctaaattctg gccgtttttg gcttttttgt
7561 tagacgaagc ttggtaccga gctcggatct ccaccccgta ccggtcctgc aggatttaaa
7621 tggcgcgccg ctagcgctac cggactcaga tctcgagctc aagcttcgaa ttctgcagtc
7681 gaattcacca tgtctagact ggacaagagc aaagtcataa acggagctct ggaattactc
7741 aatggtgtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag
7801 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc
7861 gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac
7921 tttctgcgga acaacgccaa gtcataccgc tgtgctctcc tctcacatcg cgacggggct
7981 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc
8041 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg
8101 ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa
8161 agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc
8221 gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg
8281 gagaaacagc taaagtgcga aagcggcggg ccgaccgacg cccttgacga ttttgactta
8341 gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct
8401 cttgacgatt ttgaccttga catgctcccc gggtaactaa gtaaggatcc gcggccgcac
8461 tagaggaatt ccgcccctct cccccccccc cccctaacgt tactggccga agccgcttgg
8521 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
8581 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
8641 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
8701 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
8761 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
8821 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
8881 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
8941 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
9001 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataagct taccggtacg
9061 cgtgatagct tgccacaacc cgtaccaaag atggatagat ccggaaagcc tgaactcacc
9121 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
9181 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
9241 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
9301 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg
9361 acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
9421 ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt
9481 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
9541 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
9601 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
9661 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
9721 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
9781 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
9841 tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc
9901 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
9961 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
10021 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat
10081 agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagagtag atgccgaccg
10141 aacaagagct gatttcgaga acgcctcagc cagcaactcg cgcgagccta gcaaggcaaa
10201 tgcgagagaa cggccttacg cttggtggca cagttctcgt ccacagttcg ctaagctcgc
10261 tcggctgggt cgcgggaggg ccggtcgcag tgattcaggc ccttctggat tgtgttggtc
10321 cccagggcac gattgtcatg cccacgcact cgggtgatct gactgatccc gcagattgga
10381 gatcgccgcc cgtgcctgcc gattgggtgc agatctctag atctgtacaa gtaaagcggc
10441 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
10501 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgtttagt
10561 ccctcccaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
10621 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
10681 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
10741 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
10801 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
10861 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
10921 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
10981 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
11041 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
11101 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
11161 ctttgggccg cctccccgca tcgataccgt cgacctcgat cgagacctag aaaaacatgg
11221 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca
11281 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta
11341 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
11401 tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt
11461 ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga cctttggatg
11521 gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa
11581 cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtatt
11641 agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc tgcatccgga
11701 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
11761 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
11821 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
11881 ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
11941 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
12001 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
12061 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
12121 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
12181 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
12241 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
12301 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
12361 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
12421 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
12481 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
12541 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
12601 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
12661 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
12721 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
12781 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
12841 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
12901 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
12961 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
13021 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
13081 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
13141 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
13201 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
13261 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
13321 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
13381 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
13441 caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
13501 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
13561 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
13621 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
13681 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
13741 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
13801 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
13861 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
13921 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgct
//
