| 质粒名称: | pCW57-MCS1-P2A-MCS2-mDux |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 138320 |
| 存储实验室: | Yi Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pCW57-MCS1-P2A-MCS2 (Neo) |
| 载体出品公司: | Broad Insitute |
| 空载体大小: | 7943 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,慢病毒 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Dux |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 2088 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | TRE |
| 标签/融合蛋白: | FLAG（插入件上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI(未破坏) |
| 3’克隆位点: | XhoI (未破坏) |
| 5’测序引物 | agctcgtttagtgaaccgtcagat |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCW57-MCS1-P2A-MCS2-mDux was a gift from Yi Zhang (Addgene plasmid # 138320 ; http://n2t.net/addgene:138320 ; RRID:Addgene_138320) |
| 参考文献部分: | Myc and Dnmt1 impede the pluripotent to totipotent state transition in embryonic stem cells. Fu X, Wu X, Djekidel MN, Zhang Y. Nat Cell Biol. 2019 Jul;21(7):835-844. doi: 10.1038/s41556-019-0343-0. Epub 2019 Jun 17. 10.1038/s41556-019-0343-0 PubMed 31209294 |
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-MCS1-P2A-MCS2-mDux 10031 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Dox-inducible codon-optimized mouse Dux.
ACCESSION .
VERSION .
KEYWORDS pCW57-MCS1-P2A-MCS2-mDux
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10031)
AUTHORS Fu X, Wu X, Djekidel MN, Zhang Y
TITLE Myc and Dnmt1 impede the pluripotent to totipotent state transition
in embryonic stem cells.
JOURNAL Nat Cell Biol. 2019 Jul;21(7):835-844. doi:
10.1038/s41556-019-0343-0. Epub 2019 Jun 17.
PUBMED 31209294
REFERENCE 2 (bases 1 to 10031)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10031
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 12..36
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 56..65
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 65..88
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2168..2224
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
promoter 2252..2762
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 2681..2699
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 2786..3529
/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"
CDS 3545..3598
/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 3608..4402
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3662..3681)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4272..4291
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4443..4463)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4861..4872)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5077..5310
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5382..5503
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5419..5438)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5473..5492
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 5543..5678
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5605..5624
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 5678..5749
/gene="bla"
/label=AmpR promoter
CDS 5750..6610
/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(5968..5987)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6781..7369
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7270..7289
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7523..7540
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7657..7678
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7693..7723
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7731..7747
/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 7736..7758
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7755..7771
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7755..7771
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 7790..7810
/label=T3
/note="T3 promoter, forward primer"
promoter 7792..7810
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 7838..8064
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 8065..8245
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 8292..8417
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8910..9143
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 9328..9372
/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 9585..9702
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 9756..39
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
protein_bind 9764..9782
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 9800..9818
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 9835..9853
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 9871..9889
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 9907..9925
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 9942..9960
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 9978..9996
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
1 tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagaattgg ctagcgccac
61 catggactac aaagacgatg acgacaaggg atccactagt aacggccgcc agtgtgctgg
121 aattctggct gaagctgggt ctcctgtggg aggctccgga gttgccagag agtctagaag
181 gaggaggaag actgtgtggc aagcttggca ggagcaggcc ttgttgagta ccttcaaaaa
241 gaagagatat ctctctttta aggaaaggaa ggagttggcc aaaaggatgg gggtatctga
301 ctgcaggatc agagtatggt tccaaaatag gcgaaacaga tcaggagagg aagggcatgc
361 ttcaaagagg agcataagag gatccaggcg actggcctca ccccagctcc aggaagaatt
421 ggggagtaga cctcagggca gaggcatgag atccagcggg agaaggccta gaactagact
481 gacaagtctc cagctcagga tcttggggca agcatttgag agaaatccta gaccagggtt
541 tgcaaccagg gaggagcttg ctagggacac cggcctgcca gaggacacta ttcatatatg
601 gtttcagaat agaagggctc gaagaaggca caggagggga agacctaccg cccaggatca
661 ggacctcttg gctagtcaag gatctgatgg tgcaccagcc ggcccagaag gaagagaaag
721 agagggagct caagaaaatc ttcttcccca agaagaggca ggaagcacag gaatggacac
781 atcctctcca tctgatcttc ctagcttttg tggtgaatcc caacccttcc aagtagcaca
841 accaagggga gccggacaac aagaagcacc tacaagggca gggaatgctg gatccctcga
901 accattgctg gaccagttgc ttgatgaggt gcaagttgag gaaccagcac ctgcaccact
961 gaacctggac ggagatcctg gtgggagagt acatgaaggg tcccaagaga gcttctggcc
1021 acaagaagag gcaggttcaa ctggcatgga taccagttct ccttctgatt ctaattcttt
1081 ttgtagggag agccaaccaa gccaggtggc ccaaccttgt ggtgctgggc aggaggatgc
1141 aaggactcaa gctgattcaa caggcccact tgaactgttg ttgctggacc aactgctgga
1201 tgaagttcag aaagaagagc atgtcccagt accacttgac tggggtagaa atccaggaag
1261 tagagaacac gagggcagtc aagactcact gcttcctctg gaagaggccg ttaatagtgg
1321 gatggataca agtatcccat caatatggcc tactttttgt agagagtcac agccaccaca
1381 ggtcgcacaa ccatctgggc ctggacaggc acaggcacca actcaaggag ggaatactga
1441 cccattggag ttgttccttt atcaacttct ggatgaagtg caggtggagg aacatgcacc
1501 tgctcccctc aactgggatg tggatcctgg tggaagagtg catgaggggt cttgggagag
1561 cttctggcca caggaagaag caggaagtac tggccttgat acctcatccc cttctgattc
1621 caattcattc tttagagagt ctaaaccttc tcaagtagcc caaaggagag gtgctggcca
1681 ggaggatgca aggacacagg ctgattctac cggacctctg gagctgctgc tctttgacca
1741 actgctggac gaggtgcaga aagaggaaca cgtccctgcc ccccttgact ggggaaggaa
1801 tcccggtagc atggagcatg aaggttccca ggacagcctg ctccctctgg aggaagctgc
1861 taattccggc agagatacct caatacctag tatctggccc gctttctgca gaaagtcaca
1921 acccccacaa gtggcacagc catcaggacc tggccaggca caagcaccta tacaaggagg
1981 taatacagat cccctggagt tgtttctgga tcagcttctt actgaagtac agttggagga
2041 gcagggccct gctcctgtga acgtggaaga aacatgggag caaatggaca ccacacctga
2101 tttgcccttg acctctgagg agtaccagac tcttctggac atgttgtgag tcgacaccgg
2161 ttccggagcc acgaacttct ctctgttaaa gcaagcagga gacgtggaag aaaaccccgg
2221 tcctaggctg cagacgcgtt gtacaggatc cggggttggg gttgcgcctt ttccaaggca
2281 gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg
2341 ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg
2401 ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct
2461 tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg
2521 cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata
2581 gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg
2641 ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc
2701 ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc
2761 agcaattcac ccgcgcgcgg ccaccatgtc tagactggac aagagcaaag tcataaacgg
2821 agctctggaa ttactcaatg gtgtcggtat cgaaggcctg acgacaagga aactcgctca
2881 aaagctggga gttgagcagc ctaccctgta ctggcacgtg aagaacaagc gggccctgct
2941 cgatgccctg ccaatcgaga tgctggacag gcatcatacc cacttctgcc ccctggaagg
3001 cgagtcatgg caagactttc tgcggaacaa cgccaagtca taccgctgtg ctctcctctc
3061 acatcgcgac ggggctaaag tgcatctcgg cacccgccca acagagaaac agtacgaaac
3121 cctggaaaat cagctcgcgt tcctgtgtca gcaaggcttc tccctggaga acgcactgta
3181 cgctctgtcc gccgtgggcc actttacact gggctgcgta ttggaggaac aggagcatca
3241 agtagcaaaa gaggaaagag agacacctac caccgattct atgcccccac ttctgagaca
3301 agcaattgag ctgttcgacc ggcagggagc cgaacctgcc ttccttttcg gcctggaact
3361 aatcatatgt ggcctggaga aacagctaaa gtgcgaaagc ggcgggccga ccgacgccct
3421 tgacgatttt gacttagaca tgctcccagc cgatgccctt gacgactttg accttgatat
3481 gctgcctgct gacgctcttg acgattttga ccttgacatg ctccccgggt tcgaaggaag
3541 cggagaaggt agaggttctc tcctcacttg tggtgatgtt gaagaaaacc ctggtccaat
3601 gtctagaatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
3661 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
3721 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
3781 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
3841 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
3901 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
3961 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
4021 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
4081 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
4141 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
4201 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
4261 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
4321 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
4381 ccttcttgac gagttcttct gaatgcattg aggatcgatc caagatatcg tattcttaac
4441 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
4501 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
4561 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
4621 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
4681 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
4741 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
4801 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
4861 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
4921 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
4981 gtttcgcctc ggcgtccgga ctagaggtac ctttaagacc aatgacttac aaggcagctg
5041 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
5101 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
5161 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
5221 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
5281 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
5341 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
5401 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
5461 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
5521 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
5581 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
5641 gctattccag aagtagtgag gaggcttttt tggaggcctt tcaaatatgt atccgctcat
5701 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
5761 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
5821 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
5881 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
5941 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
6001 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
6061 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
6121 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
6181 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
6241 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
6301 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
6361 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
6421 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
6481 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
6541 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
6601 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
6661 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
6721 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
6781 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
6841 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
6901 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
6961 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
7021 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
7081 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
7141 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
7201 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
7261 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
7321 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
7381 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
7441 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
7501 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
7561 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
7621 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
7681 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
7741 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
7801 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
7861 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
7921 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
7981 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
8041 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
8101 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
8161 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
8221 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
8281 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
8341 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
8401 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
8461 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
8521 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
8581 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
8641 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
8701 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
8761 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
8821 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
8881 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
8941 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
9001 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
9061 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
9121 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
9181 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
9241 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
9301 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
9361 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
9421 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
9481 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
9541 ggacaattct cgacctcgag acaaatggca gtattcatcc acaattttaa aagaaaaggg
9601 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
9661 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
9721 agcagagatc cactttggcc gcgaatcgat atgtcgagtt tactccctat cagtgataga
9781 gaacgtatgt cgagtttact ccctatcagt gatagagaac gatgtcgagt ttactcccta
9841 tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga
9901 gtttactccc tatcagtgat agagaacgta tgtcgagttt atccctatca gtgatagaga
9961 acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgaggt aggcgtgtac
10021 ggtgggaggc c
//
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