| 质粒名称: | pLV-EF1a-MinE(L4E)-EGFP_IRES-Blast |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 213117 |
| 存储实验室: | Scott Coyle |
|
载体骨架基本信息 |
|
| 载体名称: | pLV-EF1a |
| 载体出品公司: | Clontech |
| 空载体大小: | 8528bp |
| 完整质粒大小: | 9545bp |
| 载体类型: | 哺乳细胞表达, Lentiviral |
| 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: | MinE(L4E)-EGFP |
| 物种来源: | Escherichia coli |
| 插入大小: | 1017bp |
| 基因突变: | L4E |
| 启动子: | EF1a |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | EF1a_F (TCAAGCCTCAGACAGTGGTTC) |
| 3’测序引物 | EF1a_R (TAGAGGGAAACCGTTGCTAGC) |
|
文献引用方法 |
|
| 材料和方法部分: |
pLV-EF1a-MinE(L4E)-EGFP_IRES-Blast was a gift from Scott Coyle (Addgene plasmid # 213117 ; http://n2t.net/addgene:213117 ; RRID:Addgene_213117)
|
| 参考文献部分: | A programmable reaction-diffusion system for spatiotemporal cell signaling circuit design. Rajasekaran R, Chang CC, Weix EWZ, Galateo TM, Coyle SM. Cell. 2023 Dec 27:S0092-8674(23)01339-9. doi: 10.1016/j.cell.2023.12.007. 10.1016/j.cell.2023.12.007 PubMed 38181787 |
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
-EGFP_IRES-Blast质粒图谱.png)
载体简介
载体序列
LOCUS pLV-EF1a-MinE(L4E)-EGFP_IRES-Blast 9544 bp ds-DNA circular SYN 26-JAN-2024
DEFINITION Expression of a faster-running MinE mutant in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pLV-EF1a-MinE(L4E)-EGFP_IRES-Blast
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9544)
AUTHORS Rajasekaran R, Chang CC, Weix EWZ, Galateo TM, Coyle SM
TITLE A programmable reaction-diffusion system for spatiotemporal cell
signaling circuit design.
JOURNAL Cell. 2023 Dec 27:S0092-8674(23)01339-9. doi:
10.1016/j.cell.2023.12.007.
PUBMED 38181787
REFERENCE 2 (bases 1 to 9544)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 26, 2024 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9544
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 34..667
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(795..811)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(795..811)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(808..830)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 819..835
/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(843..873)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 888..909
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1026..1043)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1197..1785)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1277..1296)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1956..2816)
/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 2579..2598
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2817..2921)
/gene="bla"
/label=AmpR promoter
polyA_signal 2969..3103
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3006..3025)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3060..3079
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
LTR 3271..3904
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 3951..4076
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 4573..4806
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 4991..5035
/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 5298..5413
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
promoter 5491..5702
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
intron 5703..6641
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 6598..6618
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
regulatory 6657..6666
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 6960..7679
/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 6960..7676
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(7005..7026)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(7266..7285)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 7613..7634
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(7923..7940)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 8150..8169
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
misc_feature 8783..9371
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(8836..8856)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(9254..9265)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
ORIGIN
1 atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa
61 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga
121 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag
181 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac
241 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg
301 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact
361 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc
421 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
481 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
541 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
601 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
661 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
721 atatcagaga gtgagaggcc ttgacattgc tagcgtttac cgtcgacctc tagctagagc
781 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
841 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
901 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
961 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
1021 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
1081 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
1141 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
1201 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
1261 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
1321 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
1381 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
1441 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
1501 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
1561 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
1621 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
1681 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
1741 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
1801 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
1861 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
1921 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
1981 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
2041 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
2101 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
2161 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
2221 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
2281 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
2341 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
2401 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
2461 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
2521 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
2581 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
2641 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
2701 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
2761 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
2821 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
2881 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
2941 ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa
3001 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
3061 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc
3121 taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg
3181 gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat
3241 ttgttaaata tgtactacaa acttagtagt tggaagggct aattcactcc caaagaagac
3301 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact
3361 acacaccagg gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac
3421 cagttgagcc agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc
3481 ctgtgagcct gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca
3541 gccgcctagc atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct
3601 gatatcgagc ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg
3661 gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt
3721 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
3781 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
3841 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
3901 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
3961 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
4021 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
4081 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
4141 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
4201 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
4261 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
4321 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
4381 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg
4441 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
4501 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
4561 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
4621 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
4681 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
4741 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
4801 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
4861 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
4921 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
4981 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
5041 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
5101 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
5161 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
5221 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
5281 atctcgacgg tatcgccttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
5341 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
5401 aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttat cgatgaggcc
5461 ctttcgtctt cactcgaggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
5521 agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa
5581 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
5641 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
5701 aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg
5761 tgccttgaat tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg
5821 gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag
5881 ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct
5941 gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc
6001 tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg
6061 tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc
6121 ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg
6181 ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc
6241 ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct
6301 caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa
6361 gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca
6421 ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt
6481 tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc
6541 acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca
6601 agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggatccgcca
6661 ccatggccct tgaggacttc ttcctgtctc gaaaaaagaa cacagcgaat atcgcgaaag
6721 aacggttgca aattatagtg gcggagagac gacgaagcga cgcggagcct cattaccttc
6781 ctcaattgcg aaaagacata ctggaagtga tctgcaagta cgttcaaatt gaccctgaga
6841 tggtgaccgt gcaacttgaa cagaaggatg gagatataag catcctggaa ttgaacgtaa
6901 cccttcccga ggcggaagaa cttaaagaca gcgccggaag tgctggctct gcgggtacca
6961 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
7021 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
7081 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
7141 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
7201 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
7261 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
7321 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
7381 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
7441 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
7501 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
7561 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
7621 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag
7681 cggccgctct agagtcgacg ggccgcggta acaattgtta actaacttaa gctagcaacg
7741 gtttccctct agcgggatca attccgcccc cccccctaac gttactggcc gaagccgctt
7801 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
7861 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
7921 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
7981 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
8041 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
8101 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
8161 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
8221 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
8281 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgataatacc atggtcatga
8341 aaacatttaa catttctcaa caagatctag aattagtaga agtagcgaca gagaagatta
8401 caatgcttta tgaggataat aaacatcatg tgggagcggc aattcgtacg aaaacaggag
8461 aaatcatttc ggcagtacat attgaagcgt atataggacg agtaactgtt tgtgcagaag
8521 ccattgcgat tggtagtgca gtttcgaatg gacaaaagga ttttgacacg attgtagctg
8581 ttagacaccc ttattctgac gaagtagata gaagtattcg agtggtaagt ccttgtggta
8641 tgtgtaggga gttgatttca gactatgcac cagattgttt tgtgttaata gaaatgaatg
8701 gcaagttagt caaaactacg attgaagaac tcattccact caaatatacc cgaaattaac
8761 ccgggcgggg cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac
8821 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
8881 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
8941 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
9001 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
9061 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
9121 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
9181 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
9241 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
9301 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
9361 cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca
9421 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag
9481 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct
9541 gtag
//
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