| 质粒名称: | pCW57-CMV-ssRFP-GFP-KDEL |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 128257 |
| 存储实验室: | Noboru Mizushima |
|
载体骨架基本信息 |
|
| 载体名称: | pCW57-CMV |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7710 bp |
| 完整质粒大小: | 9213 bp |
| 载体类型: | 哺乳动物表达，慢病毒 |
| 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 |
|
| 插入基因名称: | ssRFPGFPKDEL |
| 插入大小: | 1503 bp |
| 启动子 | CMV |
| 标签/融合蛋白: | RFPGFP（主干上的N终端） |
| 克隆方法: | Gibson克隆 |
| 5' 测序引物 | CCATCCACGCTGTTTTGACCTC |
|
文献引用方法 |
|
| 材料和方法部分: | pCW57-CMV-ssRFP-GFP-KDEL was a gift from Noboru Mizushima (Addgene plasmid # 128257 ; http://n2t.net/addgene:128257 ; RRID:Addgene_128257) |
| 参考文献部分: | Intrinsically Disordered Protein TEX264 Mediates ER-phagy. Chino H, Hatta T, Natsume T, Mizushima N. Mol Cell. 2019 Apr 11. pii: S1097-2765(19)30257-6. doi: 10.1016/j.molcel.2019.03.033. 10.1016/j.molcel.2019.03.033 PubMed 31006538 |
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 pCW57-CMV-ssRFP-GFP-KDEL 9213 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Fluorescence reporter to monitor ER-phagy activity.
ACCESSION .
VERSION .
KEYWORDS pCW57-CMV-ssRFP-GFP-KDEL
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9213)
AUTHORS Chino H, Hatta T, Natsume T, Mizushima N
TITLE Intrinsically Disordered Protein TEX264 Mediates ER-phagy.
JOURNAL Mol Cell. 2019 Apr 11. pii: S1097-2765(19)30257-6. doi:
10.1016/j.molcel.2019.03.033.
PUBMED 31006538
REFERENCE 2 (bases 1 to 9213)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9213
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..227
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 228..408
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 455..580
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1073..1306
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1491..1535
/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 1724..1841
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 1909..2179
/label=tetracycline response element
/bound_moiety="tetracycline repressor TetR"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 2212..2250
/label=minimal CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2247..2271
/label=LNCX
/note="Human CMV promoter, forward primer"
sig_peptide 2350..2403
/product="Drosophila BiP signal sequence"
/label=BiP signal sequence
CDS 2404..3078
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/note="mammalian codon-optimized"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
primer_bind complement(2534..2552)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2802..2821
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 2996..3019
/label=DsRed1-C
/note="DsRed1, forward primer"
CDS 3124..3840
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 3124..3840
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3169..3190)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3430..3449)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3777..3798
/label=EGFP-C
/note="EGFP, forward primer"
promoter 3862..4372
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 4291..4309
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 4382..4978
/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(4382..4401)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4878..4898
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 4979..5032
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 5033..5779
/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"
primer_bind complement(5819..5839)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6237..6248)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6453..6686
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6758..6879
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6795..6814)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6849..6868
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 6919..7054
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6981..7000
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 7054..7125
/gene="bla"
/label=AmpR promoter
CDS 7126..7986
/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(7344..7363)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8157..8745
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8646..8665
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8899..8916
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 9033..9054
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9069..9099
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9107..9123
/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 9112..9134
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 9131..9147
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9131..9147
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 9166..9186
/label=T3
/note="T3 promoter, forward primer"
promoter 9168..9186
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
1681 agacccacct cccaaccccg aggggacaat tctcgacctc gagttttaaa agaaaagggg
1741 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
1801 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
1861 gcagagatcc actttggccg cgaatcgata tgtcctcgag tttaccactc cctatcagtg
1921 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
1981 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
2041 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
2101 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
2161 tccctatcag tgatagagaa aagtgaaagt cgagctcggt acccgggtcg aggtaggcgt
2221 gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
2281 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccgcggc
2341 cccgaattca tgaagttatg catattactg gccgtcgtgg cctttgttgg cctctcgctc
2401 gggatggcct cctccgagga cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag
2461 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
2521 ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac
2581 atcctgtccc ctcagttcca gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
2641 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
2701 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
2761 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
2821 accatgggct gggaggcctc caccgagcgg atgtaccccg aggacggcgc cctgaagggc
2881 gagatcaaga tgaggctgaa gctgaaggac ggcggccact acgacgccga ggtcaagacc
2941 acctacatgg ccaagaagcc cgtgcagctg cccggcgcct acaagaccga catcaagctg
3001 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg cgccgagggc
3061 cgccactcca ccggcgccat cgatgaattc actagtgtta acggatccct cgagcggccg
3121 cccatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
3181 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
3241 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
3301 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
3361 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
3421 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
3481 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
3541 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
3601 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
3661 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
3721 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
3781 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
3841 aaagatgagc tctaaggatc cggggttggg gttgcgcctt ttccaaggca gccctgggtt
3901 tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg
3961 gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt
4021 gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc
4081 ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg cagacggaca
4141 gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata gcggctgctc
4201 agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg ggtgtggggc
4261 ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg
4321 cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc agcaattcac
4381 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
4441 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
4501 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
4561 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
4621 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
4681 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
4741 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
4801 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
4861 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
4921 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccga
4981 aggtagaggt tctctcctca cttgtggtga tgttgaagaa aaccctggtc caatgtctag
5041 actggacaag agcaaagtca taaacggagc tctggaatta ctcaatggtg tcggtatcga
5101 aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt gagcagccta ccctgtactg
5161 gcacgtgaag aacaagcggg ccctgctcga tgccctgcca atcgagatgc tggacaggca
5221 tcatacccac ttctgccccc tggaaggcga gtcatggcaa gactttctgc ggaacaacgc
5281 caagtcatac cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc atctcggcac
5341 ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc tgtgtcagca
5401 aggcttctcc ctggagaacg cactgtacgc tctgtccgcc gtgggccact ttacactggg
5461 ctgcgtattg gaggaacagg agcatcaagt agcaaaagag gaaagagaga cacctaccac
5521 cgattctatg cccccacttc tgagacaagc aattgagctg ttcgaccggc agggagccga
5581 acctgccttc cttttcggcc tggaactaat catatgtggc ctggagaaac agctaaagtg
5641 cgaaagcggc gggccgaccg acgcccttga cgattttgac ttagacatgc tcccagccga
5701 tgcccttgac gactttgacc ttgatatgct gcctgctgac gctcttgacg attttgacct
5761 tgacatgctc cccgggtaac taagtaagga tcgatccaag atatcgtatt cttaactatg
5821 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
5881 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
5941 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
6001 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
6061 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
6121 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
6181 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
6241 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
6301 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctgttt
6361 cgcctcggcg tccggactag aggtaccttt aagaccaatg acttacaagg cagctgtaga
6421 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
6481 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
6541 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
6601 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
6661 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
6721 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
6781 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
6841 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
6901 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
6961 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
7021 ttccagaagt agtgaggagg cttttttgga ggcctttcaa atatgtatcc gctcatgaga
7081 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
7141 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
7201 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
7261 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
7321 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
7381 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
7441 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
7501 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
7561 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
7621 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
7681 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
7741 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
7801 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
7861 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
7921 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
7981 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
8041 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
8101 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
8161 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
8221 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
8281 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
8341 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
8401 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
8461 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
8521 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
8581 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
8641 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
8701 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
8761 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
8821 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
8881 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
8941 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
9001 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
9061 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
9121 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
9181 taaagggaac aaaagctgga gctgcaagct taa
//
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