| 质粒名称: | tet-pLKO-sgRNA-puro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 104321 |
| 存储实验室: | Nathanael Gray |
|
载体骨架基本信息 |
|
| 载体名称: | tet-pLKO-puro |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 8800 bp |
| 载体修饰: | shRNA编码到sgRNA编码 |
| 载体类型: | 哺乳细胞表达,慢病毒,CRISPR |
| 筛选标记: | 嘌呤霉素 |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 启动子 | H1/TO |
| 克隆方法: | 限制性内切酶 |
| 5' 测序引物 | tet-pLKO-seq-1: 5’- GTTTCAGACCCACCTCCCAAC’-3 |
|
文献引用方法 |
|
| 材料和方法部分: | tet-pLKO-sgRNA-puro was a gift from Nathanael Gray (Addgene plasmid # 104321 ; http://n2t.net/addgene:104321 ; RRID:Addgene_104321) |
| 参考文献部分: | MELK is not necessary for the proliferation of basal-like breast cancer cells. Huang HT, Seo HS, Zhang T, Wang Y, Jiang B, Li Q, Buckley DL, Nabet B, Roberts JM, Paulk J, Dastjerdi S, Winter GE, McLauchlan H, Moran J, Bradner JE, Eck MJ, Dhe-Paganon S, Zhao JJ, Gray NS. Elife. 2017 Sep 19;6. pii: e26693. doi: 10.7554/eLife.26693. 10.7554/eLife.26693 PubMed 28926338 |
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 tet-pLKO-sgRNA-puro 8880 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION 3rd generation lentiviral plasmid for inducible expression of sgRNA;
derived from tet-pLKO-puro; puromycin selection. See manual for
detailed protocols..
ACCESSION .
VERSION .
KEYWORDS tet-pLKO-sgRNA-puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8880)
AUTHORS Huang HT, Seo HS, Zhang T, Wang Y, Jiang B, Li Q, Buckley DL, Nabet
B, Roberts JM, Paulk J, Dastjerdi S, Winter GE, McLauchlan H, Moran
J, Bradner JE, Eck MJ, Dhe-Paganon S, Zhao JJ, Gray NS
TITLE MELK is not necessary for the proliferation of basal-like breast
cancer cells.
JOURNAL Elife. 2017 Sep 19;6. pii: e26693. doi: 10.7554/eLife.26693.
PUBMED 28926338
REFERENCE 2 (bases 1 to 8880)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8880
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 426..444
/label=hPGK-F
/note="Human PGK promoter, forward primer"
intron 516..1088
/label=-beta-globin intron
/note="intron from rabbit beta-globin gene"
primer_bind 1096..1115
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
promoter 1143..1161
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1172..1789
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG"
primer_bind complement(1245..1264)
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
misc_feature 1827..2400
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(1994..2011)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2221..2240
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 2420..3019
/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(2420..2439)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2916..2936
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 3147..3380
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 3452..3573
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3489..3508)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3543..3562
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 3613..3748
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3675..3694
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter complement(3769..3787)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3797..3814)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3797..3813)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3806..3828)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3955..4410
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4042..4061)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4252..4273
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 4436..4540
/gene="bla"
/label=AmpR promoter
CDS 4541..5401
/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(4759..4778)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5572..6160
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6061..6080
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6314..6331
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6448..6469
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6484..6514
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6522..6538
/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 6527..6549
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6546..6562
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6546..6562
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 6581..6601
/label=T3
/note="T3 promoter, forward primer"
promoter 6583..6601
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 6629..6855
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 6856..7036
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 7083..7208
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 7701..7934
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
primer_bind 8463..8482
/label=H1
/note="Human H1 promoter, forward primer"
protein_bind 8500..8518
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 8528..8546
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature 8711..8828
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 8877..507
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
ORIGIN
1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
481 gaccgaatca ccgacctctc tccccagggg gatctgtgag tttggggacc cttgattgtt
541 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt
601 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct
661 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa
721 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt
781 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat
841 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat
901 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacatcc
961 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga
1021 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctttt
1081 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat
1141 tgtaatacga ctcactatag ggcgaattga tatgtctaga ttagataaaa gtaaagtgat
1201 taacagcgca ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact
1261 cgcccagaag ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc
1321 tttgctcgac gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt
1381 agaaggggaa agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt
1441 actaagtcat cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta
1501 tgaaactctc gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc
1561 attatatgca ctcagcgctg tggggcattt tactttaggt tgcgtattgg aagatcaaga
1621 gcatcaagtc gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt
1681 acgacaagct atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct
1741 tgaattgatc atatgcggat tagaaaaaca acttaaatgt gaaagtgggt ccgcgtacag
1801 cggctcccgg gagttctagg gatctgcccc tctccctccc ccccccctaa cgttactggc
1861 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
1921 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
1981 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
2041 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
2101 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
2161 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
2221 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
2281 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
2341 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
2401 ggatccaccg gagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
2461 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
2521 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
2581 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
2641 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
2701 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
2761 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
2821 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
2881 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
2941 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
3001 ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc
3061 acgaccccat gcatcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
3121 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
3181 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
3241 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
3301 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
3361 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
3421 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
3481 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
3541 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
3601 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
3661 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
3721 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
3781 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
3841 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
3901 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
3961 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
4021 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
4081 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
4141 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
4201 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
4261 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
4321 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
4381 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
4441 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
4501 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
4561 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
4621 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
4681 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
4741 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
4801 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
4861 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
4921 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
4981 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
5041 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
5101 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
5161 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
5221 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
5281 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
5341 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
5401 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
5461 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
5521 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
5581 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
5641 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
5701 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
5761 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
5821 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
5881 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
5941 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
6001 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
6061 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
6121 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
6181 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
6241 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
6301 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
6361 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
6421 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
6481 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
6541 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
6601 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
6661 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
6721 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg
6781 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta
6841 gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc
6901 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
6961 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
7021 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
7081 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
7141 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
7201 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
7261 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
7321 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
7381 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
7441 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
7501 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
7561 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
7621 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
7681 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
7741 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
7801 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
7861 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
7921 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
7981 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
8041 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
8101 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
8161 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
8221 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
8281 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac
8341 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta
8401 gtgaacggat ctcgacggta tcgatcacga gactagcctc gagcggccgc aatatttgca
8461 tgtcgctatg tgttctggga aatcaccata aacgtgaaat ccctatcagt gatagagact
8521 tataagttcc ctatcagtga tagagacacc ggttctgaac agcttcttgg gaagtagttt
8581 aagagctaag ctggaaacag catagcaagt ttaaataagg ctagtccgtt atcaacttga
8641 aaaagtggca ccgagtcggt gctttttttg aattctcgac ctcgagacaa atggcagtat
8701 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
8761 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
8821 aaaattttcg ggtttattac agggacagca gagatccact ttggccgcgg ctcgaggggg
//
