| 质粒名称: | pCW-eGFP-SHLD2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 114119 |
| 存储实验室: | Daniel Durocher |
|
载体骨架基本信息 |
|
| 载体名称: | pCW57.1 |
| 载体出品公司: | Addgene #15100 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7720 bp |
| 完整质粒大小: | 11155 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒；强力霉素诱导型 |
| 筛选标记: | 嘌呤霉素 |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | SHLD2,FAM35A,RINN2 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 3435 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | TRE promoter, Tet ON |
| 标签/融合蛋白: | eGFP（插入件上的N端子） |
| 克隆方法: | Gateway克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | LNCX |
| 3' 测序引物 | O.PGK1b-R (GAACGGACGTGAAGAATGTG) |
|
文献引用方法 |
|
| 材料和方法部分: | pCW-eGFP-SHLD2 was a gift from Daniel Durocher (Addgene plasmid # 114119 ; http://n2t.net/addgene:114119 ; RRID:Addgene_114119) |
| 参考文献部分: | The shieldin complex mediates 53BP1-dependent DNA repair. Noordermeer SM, Adam S, Setiaputra D, Barazas M, Pettitt SJ, Ling AK, Olivieri M, Alvarez-Quilon A, Moatti N, Zimmermann M, Annunziato S, Krastev DB, Song F, Brandsma I, Frankum J, Brough R, Sherker A, Landry S, Szilard RK, Munro MM, McEwan A, Goullet de Rugy T, Lin ZY, Hart T, Moffat J, Gingras AC, Martin A, van Attikum H, Jonkers J, Lord CJ, Rottenberg S, Durocher D. Nature. 2018 Aug;560(7716):117-121. doi: 10.1038/s41586-018-0340-7. Epub 2018 Jul 18. 10.1038/s41586-018-0340-7 PubMed 30022168 |
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 pCW-eGFP-SHLD2 11155 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Lentiviral vector for inducible expression of N-terminally tagged
eGFP-SHLD2.
ACCESSION .
VERSION .
KEYWORDS pCW-eGFP-SHLD2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11155)
AUTHORS Noordermeer SM, Adam S, Setiaputra D, Barazas M, Pettitt SJ, Ling
AK, Olivieri M, Alvarez-Quilon A, Moatti N, Zimmermann M, Annunziato
S, Krastev DB, Song F, Brandsma I, Frankum J, Brough R, Sherker A,
Landry S, Szilard RK, Munro MM, McEwan A, Goullet de Rugy T, Lin ZY,
Hart T, Moffat J, Gingras AC, Martin A, van Attikum H, Jonkers J,
Lord CJ, Rottenberg S, Durocher D
TITLE The shieldin complex mediates 53BP1-dependent DNA repair.
JOURNAL Nature. 2018 Aug;560(7716):117-121. doi: 10.1038/s41586-018-0340-7.
Epub 2018 Jul 18.
PUBMED 30022168
REFERENCE 2 (bases 1 to 11155)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..11155
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 43..160
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 214..528
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
protein_bind 222..240
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 258..276
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 293..311
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 329..347
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 365..383
/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"
protein_bind 436..454
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 501..525
/label=LNCX
/note="Human CMV promoter, forward primer"
protein_bind 549..573
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 591..1307
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 591..1307
/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(636..657)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(897..916)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1244..1265
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind complement(4030..4054)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 4072..4089
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
promoter 4099..4609
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 4528..4546
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 4619..5215
/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(4619..4638)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 5115..5135
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 5216..5269
/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 5270..6016
/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(6056..6076)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6474..6485)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6690..6923
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6995..7116
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7032..7051)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7086..7105
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 7156..7291
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 7218..7237
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 7291..7362
/gene="bla"
/label=AmpR promoter
CDS 7363..8223
/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(7581..7600)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8394..8982
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8883..8902
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 9136..9153
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 9270..9291
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9306..9336
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9344..9360
/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 9349..9371
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 9368..9384
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9368..9384
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 9403..9423
/label=T3
/note="T3 promoter, forward primer"
promoter 9405..9423
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 9451..9677
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 9678..9858
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 9905..10030
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 10523..10756
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 10941..10985
/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 acaattctcg acctcgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
61 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
121 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
181 cagagatcca ctttggccgc gaatcgatat gtcgagttta ctccctatca gtgatagaga
241 acgtatgtcg agtttactcc ctatcagtga tagagaacga tgtcgagttt actccctatc
301 agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg tatgtcgagt
361 ttactcccta tcagtgatag agaacgtatg tcgagtttat ccctatcagt gatagagaac
421 gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgaggtag gcgtgtacgg
481 tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagaattggc
541 tagcatcaac aagtttgtac aaaaaagcag gcttcgaagg agatagaacc atggtgagca
601 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
661 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
721 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
781 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
841 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
901 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
961 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
1021 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
1081 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
1141 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
1201 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
1261 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc gcgccaagtg
1321 gaggatctca agtccacatt ttttggggtg ctccaattgc tccactgaaa atcacagtat
1381 cagaagacac agcttcttta atgtctgttg ctgacccctg gaaaaaaatt cagcttttat
1441 acagtcaaca ttctttatat ctgaaggatg aaaaacagca caaaaatctt gaaaactata
1501 aagtcccaga atctattggt tctccagatc ttagtggtca tttcttagca aactgtatga
1561 atagacatgt tcatgtgaaa gatgactttg tacgttctgt ttctgaaaca cagaatatag
1621 aatcccagaa gattcactcc tctagactga gtgatataac tagctctaat atgcaaatat
1681 gtggatttaa aagcacagtt ccgcatttca ccgaagaaga aaagtatcaa aagcttctca
1741 gtgaaaataa aattagagat gaacagccta aacatcagcc agatatatgt ggtaagaact
1801 ttaacacaaa tttgtttcag ttgggccata aatgtgcagc tgtgttggat ttggtttgta
1861 gtactgaaaa aattaatata gggcctgaag tggtacaaag agagtgtgtg ccaacagaat
1921 atcatgaaat acaaaaccag tgtttgggat tattttcctc gaacgcagta gataagtcaa
1981 ggtctgaagc agcagttagg aaggtctcag accttaaaat atcaactgat acagaatttc
2041 tcagtataat tacctccagc caggttgctt ttttagctca aaagaaagat aaaaggcgga
2101 gtcctgtaaa taaagggaat gtaaacatgg agactgaacc aaaggcaagt tacggggaga
2161 taagaatacc tgaagagaat tcgattcagc ttgatggttt tacagaagca tatgaaagtg
2221 gacaaaacca agcatattcc cttgaacttt ttagtcctgt ttgtcctaaa acagaaaata
2281 gccgcattca cataaactct gataaaggtc ttgaagaaca tacaggatct caagaacttt
2341 tcagttctga agatgaactg ccaccaaatg agatacgtat tgagttgtgt agctcaggaa
2401 tactgtgttc ccaactaaat accttccaca aaagtgctat taaaagaagc tgtacctctg
2461 aagataaagt gggccagtct gaagctctat ctagagtcct tcaagtagct aagaaaatga
2521 agttgatttc taatggagga gattctgctg tagaaatgga tcggagaaat gtgtctgaat
2581 ttaagagtat taaaaaaaca tcattaataa aaaactgtga ttctaaaagc cagaagtata
2641 attgtttagt catggtgcta tctccatgcc atgtgaagga aataaacata aaattcggac
2701 caaattctgg ctctaaagtg cctttagcaa cagttacagt aattgatcaa tcagaaacta
2761 agaagaaggt ttttctgtgg aggactgcag cattttgggc atttacagtg tttcttggag
2821 atataatttt actcacagat gttgttattc atgaggacca atggattggc gagacagtac
2881 tacaatcaac atttagcagt cagttattaa atcttgggag ttattcatct attcagcctg
2941 aagaatattc cagtgtagtt agtgaagttg tacttcaaga cttactggca tatgtgtcct
3001 caaaacattc ctacctcaga gatcttcctc cgaggcagcc tcagagggtg aacagtatag
3061 actttgtaga attggagcac cttcaacctg atgtattagt ccacgcagta ctaagagttg
3121 ttgatttcac tatactgaca gaggcagtat acagttatag aggacagaag cagaaaaaag
3181 ttatgttaac agtggaacag gcccaagatc aacattatgc gcttgtatta tggggtcctg
3241 gagcagcctg gtaccctcaa cttcaaagga aaaaaggtta tatttgggaa tttaaatatc
3301 tttttgttca gtgcaattac acactagaaa acctagaatt gcatacaacg ccttggtcat
3361 cctgtgagtg cttgtttgat gatgatataa gggcaattac atttaaagca aaatttcaaa
3421 aaagtgcacc ctcctttgtg aagatatcag acttagcaac ccacctagag gataagtgtt
3481 caggagtggt tctgattaaa gcccagattt cagagctggc atttcctatt acagcatctc
3541 agaagatagc gctaaatgct cacagttctc tgaagagtat tttttcttct cttcccaaca
3601 tcgtatatac tggttgtgca aaatgtggat tggaactaga aacagatgag aacaggatct
3661 acaaacaatg ttttagctgc ttgccattta ctatgaagaa aatatattat aggccagcgt
3721 taatgactgc cattgatgga agacatgatg tttgtatccg tgtagaatca aagctgatag
3781 agaagattct tctcaacatt tctgcagact gcctcaacag agtgatagtt ccttcctcag
3841 agatcaccta tgggatggtc gtggcagacc tgttccactc cttgttggca gtcagcgcag
3901 aaccttgtgt attaaagatt cagagccttt ttgtgttaga tgaaaacagc tatccattac
3961 aacaagattt ctccctcctg gatttttatc ctgacattgt aaagcatgga gccaatgccc
4021 gtctctagga cccagctttc ttgtacaaag tggtttagta atgaaccggt ccaccaccac
4081 caccaccact aaggatccgg ggttggggtt gcgccttttc caaggcagcc ctgggtttgc
4141 gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg accctgggtc
4201 tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta cccttgtggg
4261 ccccccggcg acgcttcctg ctccgcccct aagtcgggaa ggttccttgc ggttcgcggc
4321 gtgccggacg tgacaaacgg aagccgcacg tctcactagt accctcgcag acggacagcg
4381 ccagggagca atggcagcgc gccgaccgcg atgggctgtg gccaatagcg gctgctcagc
4441 agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt gtggggcggt
4501 agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc cggagcgcac
4561 gtcggcagtc ggctccctcg ttgaccgaat caccgacctc tctccccagc aattcaccat
4621 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
4681 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
4741 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
4801 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
4861 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
4921 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
4981 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
5041 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
5101 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
5161 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgccgaagg
5221 tagaggttct ctcctcactt gtggtgatgt tgaagaaaac cctggtccaa tgtctagact
5281 ggacaagagc aaagtcataa acggagctct ggaattactc aatggtgtcg gtatcgaagg
5341 cctgacgaca aggaaactcg ctcaaaagct gggagttgag cagcctaccc tgtactggca
5401 cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc gagatgctgg acaggcatca
5461 tacccacttc tgccccctgg aaggcgagtc atggcaagac tttctgcgga acaacgccaa
5521 gtcataccgc tgtgctctcc tctcacatcg cgacggggct aaagtgcatc tcggcacccg
5581 cccaacagag aaacagtacg aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg
5641 cttctccctg gagaacgcac tgtacgctct gtccgccgtg ggccacttta cactgggctg
5701 cgtattggag gaacaggagc atcaagtagc aaaagaggaa agagagacac ctaccaccga
5761 ttctatgccc ccacttctga gacaagcaat tgagctgttc gaccggcagg gagccgaacc
5821 tgccttcctt ttcggcctgg aactaatcat atgtggcctg gagaaacagc taaagtgcga
5881 aagcggcggg ccgaccgacg cccttgacga ttttgactta gacatgctcc cagccgatgc
5941 ccttgacgac tttgaccttg atatgctgcc tgctgacgct cttgacgatt ttgaccttga
6001 catgctcccc gggtaactaa gtaaggatcg atccaagata tcgtattctt aactatgttg
6061 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
6121 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
6181 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
6241 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
6301 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
6361 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
6421 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
6481 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
6541 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctgtttcgc
6601 ctcggcgtcc ggactagagg tacctttaag accaatgact tacaaggcag ctgtagatct
6661 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
6721 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
6781 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
6841 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
6901 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
6961 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
7021 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
7081 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
7141 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
7201 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
7261 cagaagtagt gaggaggctt ttttggaggc ctttcaaata tgtatccgct catgagacaa
7321 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
7381 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
7441 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
7501 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
7561 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
7621 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
7681 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
7741 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
7801 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
7861 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
7921 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
7981 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
8041 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
8101 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
8161 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
8221 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
8281 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
8341 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
8401 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
8461 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
8521 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
8581 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
8641 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
8701 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
8761 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
8821 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
8881 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
8941 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
9001 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
9061 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
9121 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
9181 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
9241 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
9301 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
9361 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
9421 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
9481 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
9541 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
9601 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
9661 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
9721 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
9781 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
9841 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
9901 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
9961 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
10021 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
10081 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
10141 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
10201 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
10261 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
10321 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
10381 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
10441 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
10501 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
10561 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
10621 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
10681 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
10741 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
10801 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
10861 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
10921 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
10981 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
11041 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
11101 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg agggg
//
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