| 质粒名称: | pCMV-PEmax-P2A-GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 180020 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 载体出品公司: | Clontech |
| 空载体大小: | 10578bp |
| 完整质粒大小: | 10578bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | EGFP |
| 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: | PEmax-P2A-EGFP/SV40_bpNLS-SpCas9(R221K,N394K,H840A)-SGGSx2-SV40_bpNLS-SGGSx2-MMLV_RT(D200N, T306K, W313F, T330P, L603W)-SV40_bpNLS-cMyc_NLS |
| 物种来源: | Synthetic; SpCas9 is from Streptococcus pyogenes; MMLV_RT is from the Moloney murine leukemia virus |
| 插入大小: | 7176bp |
| 启动子: | CMV |
| 标签/融合蛋白: |
SV40 bpNLS (N terminal on insert)
SV40 bpNLS (C terminal on insert)
c-Myc NLS (C terminal on insert)
P2A-EGFP (C terminal on insert)
|
| 克隆方法: | Ligation Independent Cloning |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | T7-forward |
| 3’测序引物 | BGH-reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-PEmax-P2A-GFP was a gift from David Liu (Addgene plasmid # 180020 ; http://n2t.net/addgene:180020 ; RRID:Addgene_180020) |
| 参考文献部分: | Enhanced prime editing systems by manipulating cellular determinants of editing outcomes. Chen PJ, Hussmann JA, Yan J, Knipping F, Ravisankar P, Chen PF, Chen C, Nelson JW, Newby GA, Sahin M, Osborn MJ, Weissman JS, Adamson B, Liu DR. Cell. 2021 Oct 28;184(22):5635-5652.e29. doi: 10.1016/j.cell.2021.09.018. 10.1016/j.cell.2021.09.018 PubMed 34653350 |
买家导航
pCMV-PEmax-P2A-GFP载体质粒基本信息
pCMV-PEmax-P2A-GFP质粒图谱载体图谱和pCMV-PEmax-P2A-GFP载体序列质粒序列多克隆位点信息
pCMV-PEmax-P2A-GFP质粒载体简介
pCMV-PEmax-P2A-GFP质粒序列载体序列
LOCUS pCMV-PEmax-P2A-GFP 10578 bp DNA circular SYN 14-APR-2025
DEFINITION Mammalian expression of SpCas9 PEmax prime editor with P2A-EGFP
marker.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10578)
AUTHORS Chen PJ, Hussmann JA, Yan J, Knipping F, Ravisankar P, Chen PF, Chen
C, Nelson JW, Newby GA, Sahin M, Osborn MJ, Weissman JS, Adamson B,
Liu DR
TITLE Enhanced prime editing systems by manipulating cellular determinants
of editing outcomes.
JOURNAL Cell. 2021 Oct 28;184(22):5635-5652.e29. doi:
10.1016/j.cell.2021.09.018.
PUBMED 34653350
REFERENCE 2 (bases 1 to 10578)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2021.09"; journalName: "Cell"; date: "2021-10-28-
28"; volume: "184"; issue: "22"; pages: "5635-5652.e29"
COMMENT Sequence Label: pCMV-PEmax-P2A-GFP
FEATURES Location/Qualifiers
source 1..10578
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 626..645
/label=T7
/note="T7 promoter, forward primer"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 694..714
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 715..4815
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 4873..4893
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6994..7014
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7024..7050
/codon_start=1
/product="nuclear localization signal of human c-Myc
proto-oncogene (Dang and Lee, 1988)"
/label=c-myc NLS
/translation="PAAKRVKLD"
CDS 7060..7116
/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"
CDS join(7117..7119,7120..7122,7123..7836)
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS join(7117..7119,7120..7122,7123..7833)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(7423..7442)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 7770..7791
/label=EGFP-C
/note="EGFP, forward primer"
CDS 7842..7859
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(7882..7899)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7888..8112
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(8183..8199)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8183..8199)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8196..8218)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8207..8223
/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)."
promoter complement(join(8231..8237,8238..8255,8256..8261))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8276..8297
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8414..8431)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8585..9173)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8665..8684)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(9344..10135,10136..10204))
/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 9967..9986
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10205..10309)
/gene="bla"
/label=AmpR promoter
primer_bind complement(10388..10407)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
661 aaacggacag ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtcgacaag
721 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
781 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
841 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
901 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
961 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
1021 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
1081 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
1141 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
1201 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
1261 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
1321 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag aaagctggaa
1381 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
1441 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
1501 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
1561 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
1621 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
1681 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
1741 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
1801 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
1861 gacggcaccg aggaactgct cgtgaagctg aagagagagg acctgctgcg gaagcagcgg
1921 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
1981 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
2041 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
2101 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
2161 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
2221 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
2281 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
2341 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
2401 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
2461 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
2521 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
2581 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
2641 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
2701 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
2761 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
2821 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
2881 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
2941 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
3001 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
3061 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
3121 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
3181 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacgc tatcgtgcct
3241 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
3301 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
3361 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
3421 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
3481 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
3541 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
3601 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
3661 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
3721 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
3781 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
3841 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
3901 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
3961 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
4021 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
4081 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
4141 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
4201 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
4261 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
4321 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
4381 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
4441 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
4501 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
4561 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
4621 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
4681 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
4741 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
4801 cagctgggag gtgactccgg cggaagctct ggtggcagca agcggaccgc cgacggctct
4861 gaattcgaga gccctaagaa gaaaagaaag gtgagcggag gctctagcgg cggaagcacc
4921 ctgaacattg aagacgagta tagactgcat gaaacaagca aggaacccga cgtgtccctg
4981 ggctccacct ggctgtccga ctttccccag gcctgggccg agacaggagg aatgggcctg
5041 gccgtgcggc aggcacccct gatcatccct ctgaaggcca cctctacacc cgtgagcatc
5101 aagcagtacc ctatgtctca ggaggccaga ctgggcatca agcctcacat ccagaggctg
5161 ctggaccagg gcatcctggt gccatgccag agcccctgga acacaccact gctgcccgtg
5221 aagaagccag gcaccaatga ctatagaccc gtgcaggatc tgagagaggt gaacaagagg
5281 gtggaggata tccaccccac cgtgcccaac ccttacaatc tgctgtccgg cctgccccct
5341 tctcaccagt ggtatacagt gctggacctg aaggatgcct tcttttgtct gagactgcac
5401 cctaccagcc agccactgtt cgcctttgag tggagggacc ctgagatggg catctctggc
5461 cagctgacct ggacacgcct gcctcagggc ttcaagaata gcccaacact gtttaacgag
5521 gccctgcacc gcgacctggc agatttccgg atccagcacc cagatctgat cctgctgcag
5581 tacgtggacg atctgctgct ggccgccacc agcgagctgg attgccagca gggaacacgc
5641 gccctgctgc agaccctggg aaacctggga tatagggcat ccgccaagaa ggcccagatc
5701 tgtcagaagc aggtgaagta cctgggctat ctgctgaagg agggccagag atggctgaca
5761 gaggccagga aggagacagt gatgggccag ccaacaccca agaccccaag acagctgagg
5821 gagttcctgg gcaaagcagg attttgcagg ctgttcatcc caggattcgc agagatggca
5881 gcacctctgt acccactgac caagccgggc accctgttta attggggccc tgaccagcag
5941 aaggcctatc aggagatcaa gcaggccctg ctgacagcac cagccctggg cctgccagac
6001 ctgaccaagc ctttcgagct gtttgtggat gagaagcagg gctacgccaa gggcgtgctg
6061 acccagaagc tgggaccatg gagacggccc gtggcctatc tgtccaagaa gctggaccca
6121 gtggcagcag gatggccacc atgcctgagg atggtggcag caatcgccgt gctgacaaag
6181 gatgccggca agctgaccat gggacagcca ctggtcatcc tggcaccaca cgcagtggag
6241 gccctggtga agcagcctcc agatcgctgg ctgtctaacg cccggatgac acactaccag
6301 gccctgctgc tggacaccga tcgcgtgcag tttggccctg tggtggccct gaatccagcc
6361 accctgctgc ctctgccaga ggagggcctg cagcacaact gtctggacat cctggcagag
6421 gcacacggaa caaggccaga cctgaccgat cagcccctgc ctgacgccga tcacacatgg
6481 tataccgatg gaagctccct gctgcaggag ggccagagga aggcaggagc agcagtgacc
6541 acagagacag aagtgatctg ggccaaggcc ctgccagcag gcacatccgc ccagcgggcc
6601 gagctgatcg ccctgaccca ggccctgaag atggccgagg gcaagaagct gaacgtgtac
6661 acagactcca gatatgcctt cgccaccgca cacatccacg gagagatcta caggcgccgg
6721 ggctggctga cctctgaggg caaggagatc aagaacaagg atgagatcct ggccctgctg
6781 aaggccctgt ttctgcccaa gcggctgagc atcatccact gtcctggaca ccagaaggga
6841 cactccgccg aggcaagggg caatcggatg gccgaccagg ccgccagaaa ggctgctatt
6901 actgaaactc ccgacacttc cactctgctg attgaaaact cctccccttc tggcggctca
6961 aaaagaaccg ccgacggcag cgaattcgag tctcccaaga agaagaggaa agtcggctct
7021 ggccctgccg ctaagagagt gaagctggac ggatccggcg caacaaactt ctctctgctg
7081 aaacaagccg gagatgtcga agagaatcct ggaccgatgg tgagcaaggg cgaggagctg
7141 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
7201 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
7261 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctatgga
7321 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
7381 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
7441 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
7501 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
7561 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
7621 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
7681 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
7741 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
7801 gggatcactc tcggcatgga cgagctgtac aagtaaccgg tcatcatcac catcaccatt
7861 gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
7921 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
7981 aaaatgagaa aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
8041 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
8101 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgataccg tcgacctcta
8161 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
8221 caattccaca caacatacga gccggaagca taaagtgtaa agcctagggt gcctaatgag
8281 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
8341 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
8401 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
8461 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
8521 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
8581 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
8641 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
8701 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
8761 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
8821 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
8881 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
8941 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
9001 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
9061 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
9121 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
9181 ctttgatctt ttctacgggg tctgacactc agtggaacga aaactcacgt taagggattt
9241 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
9301 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
9361 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
9421 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
9481 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
9541 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
9601 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
9661 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
9721 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
9781 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
9841 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
9901 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
9961 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
10021 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
10081 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
10141 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
10201 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
10261 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
10321 gaaaagtgcc acctgacgtc gacggatcgg gagatcgatc tcccgatccc ctagggtcga
10381 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
10441 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
10501 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
10561 gggccagata tacgcgtt
//
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