| 质粒名称: | pBK086 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 183145 |
| 存储实验室: | Daan Swarts |
|
载体骨架基本信息 |
|
| 载体名称: | pET-His6-MBP-TEV-LIC cloning vector (1M) |
| 载体出品公司: | Scott Gradia |
| 空载体大小: | 6465 bp |
| 完整质粒大小: | 9397 bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 卡那霉素 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | MapSPARTA operon (6xHis-MBP-MapTIR-APAZ and MapAgo),6xHis-MBP-MapTIR-APAZ,MapAgo |
| 物种来源: | Maribacter polysiphoniae DSM 23514 |
| 插入大小: | 4062 bp |
| GenBank编号 | WP_109649956.1 WP_109649955.1 |
| 启动子: | T7 |
| 标签/融合蛋白: |
6xHis（插件上的N端子）
MBP（插入件上的N端子）
|
| 克隆方法: | Gibson克隆 |
| 5’测序引物 | gatgaagccctgaaagacgcgcag |
| 3’测序引物 | tttgttagcagccggatctcag |
|
文献引用方法 |
|
| 材料和方法部分: | pBK086 was a gift from Daan Swarts (Addgene plasmid # 183145 ; http://n2t.net/addgene:183145 ; RRID:Addgene_183145) |
| 参考文献部分: | Short prokaryotic Argonaute systems trigger cell death upon detection of invading DNA. Koopal B, Potocnik A, Mutte SK, Aparicio-Maldonado C, Lindhoud S, Vervoort JJM, Brouns SJJ, Swarts DC. Cell, 2022. 10.1016/j.cell.2022.03.012 |
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 pBK086 9394 bp ds-DNA circular SYN 21-APR-2022
DEFINITION pET-6xHis-MBP-TEV-MapTIR-APAZ/MapAgo - Heterolgous MapSPARTA
expression.
ACCESSION .
VERSION .
KEYWORDS pBK086
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9394)
AUTHORS Koopal B, Potocnik A, Mutte SK, Aparicio-Maldonado C, Lindhoud S,
Vervoort JJM, Brouns SJJ, Swarts DC
TITLE Short prokaryotic Argonaute systems trigger cell death upon
detection of invading DNA
JOURNAL Cell, 2022.
REFERENCE 2 (bases 1 to 9394)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 21, 2022 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9394
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(278..1360)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 1322..1341
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(1361..1438)
/gene="lacI"
/label=lacI promoter
primer_bind 1644..1663
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 1747..1766
/label=T7
/note="T7 promoter, forward primer"
promoter 1747..1765
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1766..1790
/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)."
RBS 1805..1827
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1846..1863
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 1873..2973
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
primer_bind 2947..2970
/label=MBP-F
/note="Maltose binding protein, forward primer"
CDS 2986..3006
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 6004..6021
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(6074..6092)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 6088..6135
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 6172..6627
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6259..6278)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6469..6490
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(6720..7535)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 7443..7462
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 7527..7546
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 7657..8245
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8146..8165
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8399..8416
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 8431..8570
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(8496..8515)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 8656..8678
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(8672..8863)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
61 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
121 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
181 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
241 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
301 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
361 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
421 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
481 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
541 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
601 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg ataccctcat
661 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
721 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
781 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
841 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
901 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
961 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
1021 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
1081 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
1141 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
1201 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
1261 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
1321 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
1381 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg
1441 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
1501 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
1561 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
1621 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
1681 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
1741 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
1801 taattttgtt taactttaag aaggagatat accatgggtt cttctcacca tcaccatcac
1861 catggttctt ctatgaaaat cgaagaaggt aaactggtaa tctggattaa cggcgataaa
1921 ggctataacg gtctcgctga agtcggtaag aaattcgaga aagataccgg aattaaagtc
1981 accgttgagc atccggataa actggaagag aaattcccac aggttgcggc aactggcgat
2041 ggccctgaca ttatcttctg ggcacacgac cgctttggtg gctacgctca atctggcctg
2101 ttggctgaaa tcaccccgga caaagcgttc caggacaagc tgtatccgtt tacctgggat
2161 gccgtacgtt acaacggcaa gctgattgct tacccgatcg ctgttgaagc gttatcgctg
2221 atttataaca aagatctgct gccgaacccg ccaaaaacct gggaagagat cccggcgctg
2281 gataaagaac tgaaagcgaa aggtaagagc gcgctgatgt tcaacctgca agaaccgtac
2341 ttcacctggc cgctgattgc tgctgacggg ggttatgcgt tcaagtatga aaacggcaag
2401 tacgacatta aagacgtggg cgtggataac gctggcgcga aagcgggtct gaccttcctg
2461 gttgacctga ttaaaaacaa acacatgaat gcagacaccg attactccat cgcagaagct
2521 gcctttaata aaggcgaaac agcgatgacc atcaacggcc cgtgggcatg gtccaacatc
2581 gacaccagca aagtgaatta tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc
2641 aaaccgttcg ttggcgtgct gagcgcaggt attaacgccg ccagtccgaa caaagagctg
2701 gcaaaagagt tcctcgaaaa ctatctgctg actgatgaag gtctggaagc ggttaataaa
2761 gacaaaccgc tgggtgccgt agcgctgaag tcttacgagg aagagttggc gaaagatcca
2821 cgtattgccg ccactatgga aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag
2881 atgtccgctt tctggtatgc cgtgcgtact gcggtgatca acgccgccag cggtcgtcag
2941 actgtcgatg aagccctgaa agacgcgcag actaatggga tcgaggaaaa cctgtacttc
3001 caatccaatg caagaaataa aatatttata agtcacgcaa cacccgatga caatgatttt
3061 actagatggt tagcgttaaa actaattgga ttaggatacg aagtttggtg tgacattctt
3121 tttttagata aaggggttga tttctggagc aatatcgaaa aagttataag agaagatacc
3181 tgtaaatttt tacttgtttc atcatcttat tcaaatcaac gcgaaggtgt tcttaaagaa
3241 ttagcagttg cagcaaaagt aaaaaagcaa ctgaaagatg ataagttcat tattccatta
3301 gcaattgatg aacaactatc ttacgatgat attaatattg atattgttag gcttaacgct
3361 atcgatttta aaatgtcttg ggcaagaggt ttgaaagata tattagaagc gttcgaaaaa
3421 caaaaagttc caaaagaagt tgctgacgct tcaaaaagta atttacttta tcaacaaatt
3481 tttctgcacg ataaaagtgt aattgaaaaa gaagaaattt atgactcaaa ttggctttcg
3541 atattgtcct ttcctgaaga gttaagattt cacgaataca attggatgct tcctaaaaga
3601 tttgacgtaa gagaactgac gtttcctgct gttcgttaca aaaattatct ctgtacgttc
3661 gcttgggcct acgactttac atatcattta cctaaaacag aaacatatca taagagcaaa
3721 acaattagga taccaacaga ggaaatttta tcaggaagtt atgattctaa ttttattaga
3781 aatgcagaat gtaaaaggct aatagtacaa cttcttaata aagcatttga attaaggatg
3841 aaagataaag aagttcaaga atatgaaatg tctaataaaa ccgcatattg gctcgaaaaa
3901 ggtaagttag aaaaagacaa atttgagaaa accatgttag tcggaaagca aaaagataaa
3961 aattggcatt tcgcaatttc tggagcttca aaactttatc cctttcctgt attaatgatt
4021 tcatcacaca tattttttac ggcagacgga aaaaaattga ttgactcttc aagcgttcaa
4081 cattcatcaa gaagaagaca aggtaaaaac tggtggaata atacttggag aaccaaacta
4141 ttggcattta ttaaatattt atccgacgat gacacttcct tttatttaga aatgggtagt
4201 gaagagaaag tttttgtatc caatgaacca gtaaaattta aaggtaacgt aagctataat
4261 attcctgaaa aaaatactct tgaagaagaa gctgaattat cgggctttaa ccaaggcgaa
4321 gatatcgagg agctggaaga gttgattgaa aatttagaag ctgaataact cgaagcttag
4381 attttaaata acaatactaa ggaggaatta catatgaaag aacttatata catcgaagaa
4441 ccaaaaatac tatttgcaca cggtcaaaaa tgtactgacg ccagagatgg acttgcctta
4501 tttggtccat taaataatct ttatggcatt aagagtggcg ttattggtac taaacaagga
4561 ctgaaaatat ttagagatta tttagaccac atacaaaaac caatctataa tagtaacagt
4621 attactagac cgatgtttcc cggttttgaa gctgttttcg attgtaaatg ggaatccaca
4681 ggaatcacat ttaaagaagt tacaaatgaa gatattggta aatttcttta caatagcagt
4741 acgcataagc gaacttacga cttagtttcc ctttttatag acaagatcat ttctgctaat
4801 aaaaatgaag atgaaaatgt cgatgtatgg ttcgtcattg ttccagatga aatttataaa
4861 tattgcagac caaattcagt acttcccaaa gaaatggttc agactaaggc actaatgtct
4921 aaatcaaaag caaaatcttt caggtatgaa ccatcactat tcccagatat caacatagaa
4981 ttaaaagaac aagaaaaaga ggctgaaact tataattatg acgcacagtt tcacgaccaa
5041 tttaaggcaa gattattaaa acacaccata ccaacacaaa tttttagaga aagtacgttg
5101 gcttggcgcg atttcaaaaa tgcatttggt ctgccaatta gagatttttc taaaattgaa
5161 gggcatttgg cttggacaat ttcaacagct gcgttttaca aagctggagg taaaccttgg
5221 aaattatctg atgtccgaaa tggagtatgc tatcttggtt tagtttataa aaaagttgaa
5281 aaaagtaaaa acccaagaaa tgcttgttgt gcagctcaaa tgtttttaga taatggagat
5341 ggtacggttt ttaaaggaga agttggacct tggtacaatc ctaaaaatgg acagtatcat
5401 ttagaaccta aagaagcgaa ggctctgtta agtcaatcct tacaatccta taaagaacag
5461 attggagaat atcctaaaga ggtattcatt cacgcaaaaa caaggtttaa tcatcaagaa
5521 tgggatgcat ttttagaagt gacaccaaaa gaaactaatc ttgtcggagt aaccatttca
5581 aaaacaaaac cactcaaatt atacaaaact gagggagatt atacaatttt aagaggaaat
5641 gcatatgtag ttaacgagag aagtgccttt ctatggactg tgggctatgt tcctaaaata
5701 caaactgcat tatcaatgga ggttcctaat cctttattta tagaaataaa taaaggagaa
5761 gcagatataa aacaagtttt aaaggatatt ttatctttaa ctaaattgaa ttataatgcc
5821 tgcatttttg cagatggtga acctgtaacg ttaagatttg cggacaagat tggagaaata
5881 cttactgcga gtactgacat aaaaacaccg cctttagcat ttaaatatta tatctaataa
5941 cattggaagt ggataacgga tccgaattcg agcgccgtcg acaagcttgc ggccgcactc
6001 gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag
6061 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
6121 ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct
6181 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
6241 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
6301 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
6361 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
6421 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
6481 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
6541 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
6601 ttaacaaact agtaacgttt acaatttcag gtggcacttt tcggggaaat gtgcgcggaa
6661 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct
6721 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
6781 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
6841 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
6901 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
6961 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
7021 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
7081 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
7141 tgcaaccggc gcaggaacac tgccagcgca tcaacaatgt tttcacctga atcaggatat
7201 tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca
7261 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
7321 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
7381 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca
7441 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
7501 ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg
7561 taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
7621 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
7681 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
7741 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
7801 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
7861 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
7921 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
7981 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
8041 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
8101 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
8161 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
8221 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
8281 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
8341 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
8401 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
8461 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag
8521 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
8581 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
8641 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
8701 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
8761 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
8821 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
8881 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
8941 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
9001 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata
9061 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
9121 tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
9181 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
9241 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
9301 acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct
9361 tctcgccgaa acgtttggtg gcgggaccag tgac
//
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