| 质粒名称: | pLKO.1-TetON-Puro-HIF1a 0819 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 118704 |
| 存储实验室: | William Kaelin |
|
载体骨架基本信息 |
|
| 载体名称: | pLKO.1-TetON-Puro |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 8815 bp |
| 载体修饰: | -- |
| 载体类型: | 慢病毒载体 |
| 筛选标记: | 嘌呤霉素 |
| 载体抗性: | 氨苄青霉素,100μg/mL |
| 生长温度: | 30°C |
| 宿主菌: | NEB Stable |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | shRNA against human HIF1a |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | AgeI(在克隆过程中被破坏) |
| 3’克隆位点: | EcoRI(在克隆过程中被破坏) |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pLKO.1-TetON-Puro-HIF1a 0819 was a gift from William Kaelin (Addgene plasmid # 118704 ; http://n2t.net/addgene:118704 ; RRID:Addgene_118704) |
| 参考文献部分: | Paracrine Induction of HIF by Glutamate in Breast Cancer: EglN1 Senses Cysteine. Briggs KJ, Koivunen P, Cao S, Backus KM, Olenchock BA, Patel H, Zhang Q, Signoretti S, Gerfen GJ, Richardson AL, Witkiewicz AK, Cravatt BF, Clardy J, Kaelin WG Jr. Cell. 2016 Jun 30;166(1):126-39. doi: 10.1016/j.cell.2016.05.042. 10.1016/j.cell.2016.05.042 PubMed 27368101 |
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pLKO.1-TetON-Puro-HIF1a 0819载体质粒基本信息
pLKO.1-TetON-Puro-HIF1a 0819质粒图谱载体图谱和pLKO.1-TetON-Puro-HIF1a 0819载体序列质粒序列多克隆位点信息

pLKO.1-TetON-Puro-HIF1a 0819质粒载体简介
pLKO.1-TetON-Puro-HIF1a 0819质粒序列载体序列
LOCUS pLKO.1-TetON-Puro-HIF1a 0819 8815 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Lentivirus for expression of shRNA1 against human HIF1a
(Dox-inducible) .
ACCESSION .
VERSION .
KEYWORDS pLKO.1-TetON-Puro-HIF1a 0819
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8815)
AUTHORS Briggs KJ, Koivunen P, Cao S, Backus KM, Olenchock BA, Patel H,
Zhang Q, Signoretti S, Gerfen GJ, Richardson AL, Witkiewicz AK,
Cravatt BF, Clardy J, Kaelin WG Jr
TITLE Paracrine Induction of HIF by Glutamate in Breast Cancer: EglN1
Senses Cysteine.
JOURNAL Cell. 2016 Jun 30;166(1):126-39. doi: 10.1016/j.cell.2016.05.042.
PUBMED 27368101
REFERENCE 2 (bases 1 to 8815)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8815
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 324..418
/label=H1-2O2 promoter
/note="doxycycline-inducible variant of the human H1 RNA
promoter (Henriksen et al., 2007)"
primer_bind 335..354
/label=H1
/note="Human H1 promoter, forward primer"
protein_bind 372..390
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 400..418
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature 518..635
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 684..1194
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 1113..1131
/label=hPGK-F
/note="Human PGK promoter, forward primer"
intron 1203..1775
/label=-beta-globin intron
/note="intron from rabbit beta-globin gene"
primer_bind 1783..1802
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
promoter 1830..1848
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1859..2476
/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(1932..1951)
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
misc_feature 2514..3087
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2681..2698)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2908..2927
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3107..3706
/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(3107..3126)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3603..3623
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 3834..4067
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4139..4260
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4176..4195)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4230..4249
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4300..4435
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4362..4381
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter complement(4456..4474)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4484..4501)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4484..4500)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4493..4515)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4642..5097
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4729..4748)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4939..4960
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5123..5227
/gene="bla"
/label=AmpR promoter
CDS 5228..6088
/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(5446..5465)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6259..6847
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6748..6767
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7001..7018
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7135..7156
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7171..7201
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7209..7225
/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 7214..7236
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7233..7249
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7233..7249
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 7268..7288
/label=T3
/note="T3 promoter, forward primer"
promoter 7270..7288
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 7316..7542
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 7543..7723
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 7770..7895
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8388..8621
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 8806..35
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
ORIGIN
1 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
61 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
121 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
181 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
241 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgatcac
301 gagactagcc tcgagcggcc gcaatatttg catgtcgcta tgtgttctgg gaaatcacca
361 taaacgtgaa atccctatca gtgatagaga cttataagtt ccctatcagt gatagagaca
421 ccggtgctct ttgtggttgg atctactcga gtagatccaa ccacaaagag catttttaat
481 tctcgacctc gagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
541 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
601 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
661 atccactttg gccgcggctc gagggggttg gggttgcgcc ttttccaagg cagccctggg
721 tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct
781 gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt
841 gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc
901 gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga
961 cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc
1021 tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg
1081 gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa gcctccggag
1141 cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc ccagggggat
1201 ctgtgagttt ggggaccctt gattgttctt tctttttcgc tattgtaaaa ttcatgttat
1261 atggaggggg caaagttttc agggtgttgt ttagaatggg aagatgtccc ttgtatcacc
1321 atggaccctc atgataattt tgtttctttc actttctact ctgttgacaa ccattgtctc
1381 ctcttatttt cttttcattt tctgtaactt tttcgttaaa ctttagcttg catttgtaac
1441 gaatttttaa attcactttt gtttatttgt cagattgtaa gtactttctc taatcacttt
1501 tttttcaagg caatcagggt atattatatt gtacttcagc acagttttag agaacaattg
1561 ttataattaa atgataaggt agaatatttc tgcatataaa ttctggctgg cgtggaaata
1621 ttcttattgg tagaaacaac tacatcctgg tcatcatcct gcctttctct ttatggttac
1681 aatgatatac actgtttgag atgaggataa aatactctga gtccaaaccg ggcccctctg
1741 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
1801 tgctgtctca tcattttggc aaagaattgt aatacgactc actatagggc gaattgatat
1861 gtctagatta gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg
1921 aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt
1981 gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga
2041 taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa
2101 taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt
2161 aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg
2221 ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgctgtgg ggcattttac
2281 tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc
2341 tactactgat agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg
2401 tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact
2461 taaatgtgaa agtgggtccg cgtacagcgg ctcccgggag ttctagggat ctgcccctct
2521 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
2581 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
2641 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
2701 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
2761 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
2821 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
2881 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
2941 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
3001 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
3061 gttttccttt gaaaaacacg atgataagga tccaccggag cttaccatga ccgagtacaa
3121 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
3181 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
3241 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
3301 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
3361 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
3421 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
3481 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
3541 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
3601 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
3661 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga
3721 cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcggtacctt taagaccaat
3781 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
3841 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
3901 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
3961 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
4021 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
4081 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
4141 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
4201 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
4261 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
4321 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
4381 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg
4441 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
4501 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
4561 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
4621 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
4681 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
4741 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
4801 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
4861 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
4921 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
4981 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
5041 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
5101 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
5161 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
5221 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
5281 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
5341 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
5401 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
5461 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
5521 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
5581 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
5641 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
5701 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
5761 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
5821 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
5881 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
5941 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
6001 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
6061 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
6121 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
6181 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
6241 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
6301 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
6361 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
6421 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
6481 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
6541 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
6601 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
6661 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
6721 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
6781 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
6841 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
6901 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
6961 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
7021 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
7081 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
7141 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
7201 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
7261 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag
7321 tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta
7381 caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc
7441 ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat
7501 tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac
7561 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
7621 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
7681 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
7741 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
7801 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
7861 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
7921 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
7981 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
8041 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
8101 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
8161 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
8221 ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
8281 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
8341 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
8401 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
8461 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
8521 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
8581 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
8641 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
8701 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
8761 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaac
//
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