| 质粒名称: | pSMPP-mCherry-mTrim21 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 104971 |
| 存储实验室: | Leo James |
|
载体骨架基本信息 |
|
| 载体名称: | pSMPP |
| 载体出品公司: | William A.McEwan（MRC LMB） |
| 空载体大小: | 10016 bp |
| 完整质粒大小: | 12127 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: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | Tripartite Motif containing 21 |
| 物种来源: | M.musculus（小鼠） |
| 插入大小: | 1428 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | SFFV |
| 标签/融合蛋白: | mCherry（插入件上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NotI(未破坏) |
| 3’克隆位点: | NdeI (未破坏) |
| 5’测序引物 | TGCTTCTCGCTTCTGTTCG |
| 3’测序引物 | ATAGCGTAAAAGGAGCAACATAG |
|
文献引用方法 |
|
| 材料和方法部分: | pSMPP-mCherry-mTrim21 was a gift from Leo James (Addgene plasmid # 104971 ; http://n2t.net/addgene:104971 ; RRID:Addgene_104971) |
| 参考文献部分: | A Method for the Acute and Rapid Degradation of Endogenous Proteins. Clift D, McEwan WA, Labzin LI, Konieczny V, Mogessie B, James LC, Schuh M. Cell. 2017 Nov 13. pii: S0092-8674(17)31255-2. doi: 10.1016/j.cell.2017.10.033. 10.1016/j.cell.2017.10.033 PubMed 29153837 |
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 pSMPP-mCherry-mTrim21 12130 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Lentiviral Transfer Plasmid with mCherry-tagged mouse Trim21.
ACCESSION .
VERSION .
KEYWORDS pSMPP-mCherry-mTrim21
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12130)
AUTHORS Clift D, McEwan WA, Labzin LI, Konieczny V, Mogessie B, James LC,
Schuh M
TITLE A Method for the Acute and Rapid Degradation of Endogenous Proteins.
JOURNAL Cell. 2017 Nov 13. pii: S0092-8674(17)31255-2. doi:
10.1016/j.cell.2017.10.033.
PUBMED 29153837
REFERENCE 2 (bases 1 to 12130)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..12130
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 91..724
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 771..896
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1393..1626
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1811..1855
/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"
misc_feature 2140..2257
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2410..2817
/label=SFFV promoter
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
regulatory 2856..2865
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2862..3569
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(3007..3025)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3275..3294
/label=mCherry-F
/note="mCherry, forward primer"
CDS 3606..4994
/codon_start=1
/product="mouse RING E3 ubiquitin ligase that can be used
together with antibodies to degrade intracellular proteins
(Clift et al., 2018)"
/label=TRIM21
/note="binds to the Fc domain of antibodies"
/translation="MSLEKMWEEVTCSICLDPMVEPMSIECGHCFCKECIFEVGKNGGS
SCPECRQQFLLRNLRPNRHIANMVENLKQIAQNTKKSTQETHCMKHGEKLHLFCEEDGQ
ALCWVCAQSGKHRDHTRVPIEEAAKVYQEKIHVALEKLRKGKELAEKMEMDLTMQRTDW
KRNIDTQKSRIHAEFALQNSLLAQEEQRQLQRLEKDQREYLRLLGKKEAELAEKNQALQ
ELISELERRIRGSELELLQEVRIILERSGSWNLDTLDIDAPDLTSTCPVPGRKKMLRTC
WVHITLDRNTANSWLIISKDRRQVRMGDTHQNVSDNKERFSNYPMVLGAQRFSSGKMYW
EVDVTQKEAWDLGVCRDSVQRKGQFSLSPENGFWTIWLWQDSYEAGTSPQTTLHIQVPP
CQIGIFVDYEAGVVSFYNITDHGSLIYTFSECVFAGPLRPFFNVGFNYSGGNAAPLKLC
PLKM"
primer_bind complement(5107..5127)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5525..5536)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
promoter 5568..6067
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(5603..5626)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 6000..6019
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 6088..6687
/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(6088..6107)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 6584..6604
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 6782..7015
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(7040..7058)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(7041..7058)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(7348..7367)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(7581..7600)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7700..7722
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(7760..7778)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7846..7950
/gene="bla"
/label=AmpR promoter
CDS 7951..8811
/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(8169..8188)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8982..9570
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9471..9490
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 9724..9741
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(9811..9831)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 9816..10145
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 9996..10131
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 10058..10077
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
intron 11279..11344
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 11474..11494
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 11919..12053
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(11956..11975)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 12010..12029
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat
61 ttgttaaata tgtactacaa acttagtagt tggaagggct aattcactcc caaagaagac
121 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact
181 acacaccagg gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac
241 cagttgagcc agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc
301 ctgtgagcct gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca
361 gccgcctagc atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct
421 gatatcgagc ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg
481 gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt
541 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
601 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
661 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
721 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
781 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
841 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
901 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
961 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
1021 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
1081 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
1141 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
1201 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg
1261 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
1321 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
1381 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
1441 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
1501 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
1561 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
1621 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
1681 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
1741 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
1801 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
1861 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
1921 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
1981 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
2041 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
2101 atctcgacgg tatcgccaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
2161 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
2221 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
2281 agatccagtt tggatcgata agcttgatat cgaattcctg cagccccgat aaaataaaag
2341 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
2401 ctagctgcag taacgccatt ttgcaaggca tggaaaaata ccaaaccaag aatagagaag
2461 ttcagatcaa gggcgggtac atgaaaatag ctaacgttgg gccaaacagg atatctgcgg
2521 tgagcagttt cggccccggc ccggggccaa gaacagatgg tcaccgcagt ttcggccccg
2581 gcccgaggcc aagaacagat ggtccccaga tatggcccaa ccctcagcag tttcttaaga
2641 cccatcagat gtttccaggc tcccccaagg acctgaaatg accctgcgcc ttatttgaat
2701 taaccaatca gcctgcttct cgcttctgtt cgcgcgcttc tgcttcccga gctctataaa
2761 agagctcaca acccctcact cggcgcgcca gtcctccgac agactgagtc gcccgggggg
2821 gatccactgt gatcatcaga cgcgtacagc atatggccac catggtgagc aagggcgagg
2881 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
2941 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
3001 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
3061 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
3121 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
3181 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
3241 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
3301 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
3361 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
3421 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
3481 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
3541 ccaccggcgg catggacgag ctgtacaagt ccggactcag atctcgagct caagcttcga
3601 attcaatgtc tctggaaaag atgtgggagg aggtcacctg ttctatctgc ctggatccca
3661 tggtggagcc tatgagtatc gaatgtggcc attgcttttg caaggaatgc atttttgaag
3721 ttgggaagaa tgggggcagt tcatgtcccg agtgccggca acagtttctg ctccgaaacc
3781 tcaggcccaa tagacatata gccaacatgg tggaaaacct taaacagata gcccagaata
3841 ccaagaagag tacccaggaa acgcactgca tgaagcatgg agagaagctt cacctattct
3901 gtgaggaaga tgggcaggcc ctttgctggg tgtgtgccca gtctgggaaa caccgggacc
3961 acaccagggt ccctattgaa gaggctgcta aggtatacca ggagaagatc cacgtggctt
4021 tagaaaaact gagaaagggg aaagagttgg ccgagaagat ggaaatggat ctcacgatgc
4081 aaagaacaga ctggaagagg aacattgaca cccagaagtc gaggattcac gcagagttcg
4141 cacttcagaa tagcttgctg gctcaggagg agcagaggca gctgcagagg ctggagaagg
4201 atcaaaggga gtacctgaga ctcctgggga agaaggaggc tgagctggct gagaagaacc
4261 aggccctgca ggagctgatc tcagagctgg agaggaggat tcgtggttca gagctggagc
4321 tactgcagga ggtgaggatc atcctggaaa ggagtggatc ctggaacctg gacacgttag
4381 atattgacgc cccagaccta acaagcacat gccctgtgcc agggcggaag aagatgctga
4441 ggacgtgttg ggttcatatt actctggatc gcaacaccgc caactcatgg ctcatcatct
4501 caaaggatcg gagacaagtg aggatgggag acacccatca gaacgtgtct gacaataagg
4561 agaggtttag taattacccc atggtgctag gtgcccagag attctcctct gggaagatgt
4621 actgggaggt agatgtgact caaaaggagg cctgggatct gggggtttgc agagattctg
4681 ttcagaggaa agggcagttt tcactcagtc ccgagaatgg cttctggacc atttggttat
4741 ggcaagacag ctatgaggct ggtaccagtc ctcagaccac cctccacatt caagtacctc
4801 catgccaaat tgggatcttt gtggactatg aggctggcgt tgtctccttc tacaacataa
4861 ctgaccatgg ctccctcatt tacaccttct cggagtgtgt ttttgctgga cctctgcgac
4921 ctttcttcaa tgttggtttc aattatagtg ggggaaatgc agcgcctcta aagctctgtc
4981 cactaaagat gtgagcggcc gcgactctag agtcgacctg caggcatgca agcttgatat
5041 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
5101 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
5161 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
5221 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
5281 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
5341 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
5401 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
5461 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
5521 cccttcggcc ctcaatccag cggaccttcc ttcccatcaa ttctaccggg taggggaggc
5581 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
5641 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
5701 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
5761 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
5821 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
5881 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
5941 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
6001 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
6061 cctttcgacc tgcagcccaa gcttaccatg accgagtaca agcccacggt gcgcctcgcc
6121 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
6181 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
6241 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
6301 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
6361 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
6421 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
6481 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
6541 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
6601 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
6661 tgcatgaccc gcaagcccgg tgcctgactc gagggaatta attcgagctc ggtaccttta
6721 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
6781 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
6841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
6901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
6961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagcatc
7021 tagaattaat tccgtgtatt ctatagtgtc acctaaatcg tatgtgtatg atacataagg
7081 ttatgtatta attgtagccg cgttctaacg acaatatgta caagcctaat tgtgtagcat
7141 ctggcttact gaagcagacc ctatcatctc tctcgtaaac tgccgtcaga gtcggtttgg
7201 ttggacgaac cttctgagtt tctggtaacg ccgtcccgca cccggaaatg gtcagcgaac
7261 caatcagcag ggtcatcgct agccagatcc tctacgccgg acgcatcgtg gccggcatca
7321 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
7381 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
7441 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc
7501 tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc
7561 gtcgaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
7621 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
7681 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
7741 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
7801 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
7861 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
7921 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
7981 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
8041 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
8101 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
8161 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
8221 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
8281 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
8341 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
8401 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
8461 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
8521 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
8581 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
8641 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
8701 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
8761 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
8821 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
8881 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
8941 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
9001 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
9061 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
9121 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
9181 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
9241 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
9301 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
9361 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
9421 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
9481 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
9541 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
9601 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
9661 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
9721 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
9781 gcgcgttggc cgattcatta atgcagctgt ggaatgtgtg tcagttaggg tgtggaaagt
9841 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
9901 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
9961 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
10021 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
10081 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
10141 gcaaaaagct tggacacaag acaggcttgc gagatatgtt tgagaatacc actttatccc
10201 gcgtcaggga gaggcagtgc gtaaaaagac gcggactcat gtgaaatact ggtttttagt
10261 gcgccagatc tctataatct cgcgcaacct attttcccct cgaacacttt ttaagccgta
10321 gataaacagg ctgggacact tcacatgagc gaaaaataca tcgtcacctg ggacatgttg
10381 cagatccatg cacgtaaact cgcaagccga ctgatgcctt ctgaacaatg gaaaggcatt
10441 attgccgtaa gccgtggcgg tctgtaccgg gtgcgttact ggcgcgtgaa ctgggtattc
10501 gtcatgtcga taccgtttgt atttccagct acgatcacga caaccagcgc gagcttaaag
10561 tgctgaaacg cgcagaaggc gatggcgaag gcttcatcgt tattgatgac ctggtggata
10621 ccggtggtac tgcggttgcg attcgtgaaa tgtatccaaa agcgcacttt gtcaccatct
10681 tcgcaaaacc ggctggtcgt ccgctggttg atgactatgt tgttgatatc ccgcaagata
10741 cctggattga acagccgtgg gatatgggcg tcgtattcgt cccgccaatc tccggtcgct
10801 aatcttttca acgcctggca ctgccgggcg ttgttctttt taacttcagg cgggttacaa
10861 tagtttccag taagtattct ggaggctgca tccatgacac aggcaaacct gagcgaaacc
10921 ctgttcaaac cccgctttaa acatcctgaa acctcgacgc tagtccgccg ctttaatcac
10981 ggcgcacaac cgcctgtgca gtcggccctt gatggtaaaa ccatccctca ctggtatcgc
11041 atgattaacc gtctgatgtg gatctggcgc ggcattgacc cacgcgaaat cctcgacgtc
11101 caggcacgta ttgtgatgag cgatgccgaa cgtaccgacg atgatttata cgatacggtg
11161 attggctacc gtggcggcaa ctggatttat gagtgggccc cggatctttg tgaaggaacc
11221 ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa gctctaaggt
11281 aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg tttgtgtatt
11341 ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct ttaatgagga
11401 aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg ctgactctca
11461 acattctact cctccaaaaa agaagagaaa ggtagaagac cccaaggact ttccttcaga
11521 attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt gctttgctat
11581 ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa aatattctgt
11641 aacctttata agtaggcata acagttataa tcataacata ctgttttttc ttactccaca
11701 caggcataga gtgtctgcta ttaataacta tgctcaaaaa ttgtgtacct ttagcttttt
11761 aatttgtaaa ggggttaata aggaatattt gatgtatagt gccttgacta gagatcataa
11821 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
11881 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
11941 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
12001 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga
12061 taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca
12121 attacctgtg
//
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