| 质粒名称: | pEvolvR-enCas9-PolI3M-TBD |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 113077 |
| 存储实验室: | John Dueber, David Schaffer |
|
载体骨架基本信息 |
|
| 载体名称: | -- |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | enCas9-PolI3M-TBD |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pEvolvR-enCas9-PolI3M-TBD was a gift from John Dueber & David Schaffer (Addgene plasmid # 113077 ; http://n2t.net/addgene:113077 ; RRID:Addgene_113077) |
| 参考文献部分: | CRISPR-guided DNA polymerases enable diversification of all nucleotides in a tunable window. Halperin SO, Tou CJ, Wong EB, Modavi C, Schaffer DV, Dueber JE. Nature. 2018 Aug 1. pii: 10.1038/s41586-018-0384-8. doi: 10.1038/s41586-018-0384-8. 10.1038/s41586-018-0384-8 PubMed 30069054 |
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 pEvolvR-enCas9-PolI3M-TBD 12753 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses enCas9 fused to PolI3M-TBD in E. coli. contains a
BsmbI-flanked GFP cassette for sgRNA cloning..
ACCESSION .
VERSION .
KEYWORDS pEvolvR-enCas9-PolI3M-TBD
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12753)
AUTHORS Halperin SO, Tou CJ, Wong EB, Modavi C, Schaffer DV, Dueber JE
TITLE CRISPR-guided DNA polymerases enable diversification of all
nucleotides in a tunable window.
JOURNAL Nature. 2018 Aug 1. pii: 10.1038/s41586-018-0384-8. doi:
10.1038/s41586-018-0384-8.
PUBMED 30069054
REFERENCE 2 (bases 1 to 12753)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..12753
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 23..78
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 59..77
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
RBS 96..107
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
RBS 5000..5008
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS complement(7417..8232)
/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 8140..8159
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 8224..8243
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 8461..9049
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8950..8969
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 9203..9220
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 9235..9374
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(9300..9319)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 9460..9482
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(9476..9667)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
RBS 9797..9808
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 9815..10474
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKF*PAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(9881..9900)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
RBS 10774..10785
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 10792..11505
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
terminator 11517..11588
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 11604..11631
/label=T7Te terminator
/note="phage T7 early transcription terminator"
misc_RNA 11645..11720
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 11741..11770
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 11786..11803
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(11859..11876)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 12029..12075
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(12134..4)
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind 12665..12684
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
ORIGIN
1 acatcattaa ttcctaattt ttgttgacac tctatcgttg atagagttat tttaccactc
61 cctatcagtg atagagaaaa gaattcaaaa gatctaaaga ggagaaaaga tctatggaca
121 agaagtattc tatcggactg gctatcggga ctaatagcgt cgggtgggcc gtgatcactg
181 acgagtacaa ggtgccctct aagaagttca aggtgctcgg gaacaccgac cggcattcca
241 tcaagaaaaa tctgatcgga gctctcctct ttgattcagg ggaaaccgct gaagcaaccc
301 gcctcaagcg gactgctaga cggcggtaca ccaggaggaa gaaccggatt tgttaccttc
361 aagagatatt ctccaacgaa atggcaaagg tcgacgacag cttcttccat aggctggaag
421 aatcattcct cgtggaagag gataagaagc atgaacggca tcccatcttc ggtaatatcg
481 tcgacgaggt ggcctatcac gagaaatacc caaccatcta ccatcttcgc aaaaagctgg
541 tggactcaac cgacaaggca gacctccggc ttatctacct ggccctggcc cacatgatca
601 agttcagagg ccacttcctg atcgagggcg acctcaatcc tgacaatagc gatgtggata
661 aactgttcat ccagctggtg cagacttaca accagctctt tgaagagaac cccatcaatg
721 caagcggagt cgatgccaag gccattctgt cagcccggct gtcaaagagc cgcagacttg
781 agaatcttat cgctcagctg ccgggtgaaa agaaaaatgg actgttcggg aacctgattg
841 ctctttcact tgggctgact cccaatttca agtctaattt cgacctggca gaggatgcca
901 agctgcaact gtccaaggac acctatgatg acgatctcga caacctcctg gcccagatcg
961 gtgaccaata cgccgacctt ttccttgctg ctaagaatct ttctgacgcc atcctgctgt
1021 ctgacattct ccgcgtgaac actgaaatca ccaaggcccc tctttcagct tcaatgatta
1081 agcggtatga tgagcaccac caggacctga ccctgcttaa ggcactcgtc cggcagcagc
1141 ttccggagaa gtacaaggaa atcttctttg accagtcaaa gaatggatac gccggctaca
1201 tcgacggagg tgcctcccaa gaggaatttt ataagtttat caaacctatc cttgagaaga
1261 tggacggcac cgaagagctc ctcgtgaaac tgaatcggga ggatctgctg cggaagcagc
1321 gcactttcga caatgggagc attccccacc agatccatct tggggagctt cacgccatcc
1381 ttcggcgcca agaggacttc tacccctttc ttaaggacaa cagggagaag attgagaaaa
1441 ttctcacttt ccgcatcccc tactacgtgg gacccctcgc cagaggaaat agccggtttg
1501 cttggatgac cagaaagtca gaagaaacta tcactccctg gaacttcgaa gaggtggtgg
1561 acaagggagc cagcgctcag tcattcatcg aacggatgac taacttcgat aagaacctcc
1621 ccaatgagaa ggtcctgccg aaacattccc tgctctacga gtactttacc gtgtacaacg
1681 agctgaccaa ggtgaaatat gtcaccgaag ggatgaggaa gcccgcattc ctgtcaggcg
1741 aacaaaagaa ggcaattgtg gaccttctgt tcaagaccaa tagaaaggtg accgtgaagc
1801 agctgaagga ggactatttc aagaaaattg aatgcttcga ctctgtggag attagcgggg
1861 tcgaagatcg gttcaacgca agcctgggta cttaccatga tctgcttaag atcatcaagg
1921 acaaggattt tctggacaat gaggagaacg aggacatcct tgaggacatt gtcctgactc
1981 tcactctgtt cgaggaccgg gaaatgatcg aggagaggct taagacctac gcccatctgt
2041 tcgacgataa agtgatgaag caacttaaac ggagaagata taccggatgg ggacgcctta
2101 gccgcaaact catcaacgga atccgggaca aacagagcgg aaagaccatt cttgatttcc
2161 ttaagagcga cggattcgct aatcgcaact tcatgcaact tatccatgat gattccctga
2221 cctttaagga ggacatccag aaggcccaag tgtctggaca aggtgactca ctgcacgagc
2281 atatcgcaaa tctggctggt tcacccgcta ttaagaaggg tattctccag accgtgaaag
2341 tcgtggacga gctggtcaag gtgatgggtc gccataaacc agagaacatt gtcatcgaga
2401 tggccaggga aaaccagact acccagaagg gacagaagaa cagcagggag cggatgaaaa
2461 gaattgagga agggattaag gagctcgggt cacagatcct taaagagcac ccggtggaaa
2521 acacccagct tcagaatgag aagctctatc tgtactacct tcaaaatgga cgcgatatgt
2581 atgtggacca agagcttgat atcaacaggc tctcagacta cgacgtggac catatcgtcc
2641 ctcagagctt cctcgcagac gactcaattg acaataaggt gctgactcgc tcagacaaga
2701 accggggaaa gtcagataac gtgccctcag aggaagtcgt gaaaaagatg aagaactatt
2761 ggcgccagct tctgaacgca aagctgatca ctcagcggaa gttcgacaat ctcactaagg
2821 ctgagagggg cggactgagc gaactggaca aagcaggatt cattaaacgg caacttgtgg
2881 agactcggca gattactaaa catgtcgccc aaatccttga ctcacgcatg aataccaagt
2941 acgacgaaaa cgacaaactt atccgcgagg tgaaggtgat taccctgaag tccaagctgg
3001 tcagcgattt cagaaaggac tttcaattct acaaagtgcg ggagatcaat aactatcatc
3061 atgctcatga cgcatatctg aatgccgtgg tgggaaccgc cctgatcaag aagtacccag
3121 cactggaaag cgagttcgtg tacggagact acaaggtcta cgacgtgcgc aagatgattg
3181 ccaaatctga gcaggagatc ggaaaggcca ccgcaaagta cttcttctac agcaacatca
3241 tgaatttctt caagaccgaa atcacccttg caaacggtga gatccggaag gcgccgctca
3301 tcgagactaa tggggagact ggcgaaatcg tgtgggacaa gggcagagat ttcgctaccg
3361 tgcgcaaagt gctttctatg cctcaagtga acatcgtgaa gaaaaccgag gtgcaaaccg
3421 gaggcttttc taaggaatca atcctcccca agcgcaactc cgacaagctc attgcaagga
3481 agaaggattg ggaccctaag aagtacggcg gattcgattc accaactgtg gcttattctg
3541 tcctggtcgt ggctaaggtg gaaaaaggaa agtctaagaa gctcaagagc gtgaaggaac
3601 tgctgggtat caccattatg gagcgcagct ccttcgagaa gaacccaatt gactttctcg
3661 aagccaaagg ttacaaggaa gtcaagaagg accttatcat caagctccca aagtatagcc
3721 tgttcgaact ggagaatggg cggaagcgga tgctcgcctc cgctggcgaa cttcagaagg
3781 gtaatgagct ggctctcccc tccaagtacg tgaatttcct ctaccttgca agccattacg
3841 agaagctgaa ggggagcccc gaggacaacg agcaaaagca actgtttgtg gagcagcata
3901 agcattatct ggacgagatc attgagcaga tttccgagtt ttctaaacgc gtcattctcg
3961 ctgatgccaa cctcgataaa gtccttagcg catacaataa gcacagagac aaaccaattc
4021 gggagcaggc tgagaatatc atccacctgt tcaccctcac caatcttggt gcccctgccg
4081 cattcaagta cttcgacacc accatcgacc ggaaacgcta tacctccacc aaagaagtgc
4141 tggacgccac cctcatccac cagagcatca ccggacttta cgaaactcgg attgacctct
4201 cacagctcgg aggggatggt tctagtgaaa ccccgggaac aagtgagtcg gccacccctg
4261 aaggtggatc agggggtagc ggatccgttc agatcccgca gaacccgctg attctggttg
4321 acggatctag ttacctgtac cgtgcttacc atgctttccc gcctttgacc aattctgctg
4381 gtgaacctac gggagctatg tacggagttc tgaatatgtt gcgttcttta attatgcagt
4441 acaagcctac ccacgctgct gttgttttcg atgctaaagg taagacgttc cgcgacgagt
4501 tattcgagca ctataagtct caccgtcctc cgatgcctga tgacttacgc gctcagattg
4561 agccgctgca tgctatggtg aaggctatgg gtttacctct tttggctgtc agcggtgttg
4621 aggctgatga tgtcattggc accttagctc gtgaggctga gaaggctggt cgccctgttt
4681 tgatttctac cggtgacaag gacatggctc aattggttac cccgaacatc accctgatca
4741 acaccatgac caacacgatt ctgggtcctg aggaagttgt taacaaatat ggtgttcctc
4801 cggagttgat tattgacttt cttgctctta tgggcgattc ttcagacaat atcccgggtg
4861 ttccaggtgt tggagagaag actgctcaag ctctgcttca gggtctgggt ggtttggaca
4921 ccctttacgc tgaaccggag aagatcgccg gtctgtcttt tcgcggtgct aagaccatgg
4981 ctgctaaact ggaacagaat aaggaggtcg catacctgtc ttatcaattg gctaccatca
5041 agacggatgt ggagttagaa cttacgtgcg agcagcttga ggttcaacag cctgctgctg
5101 aggaactgct gggtcttttt aagaaatacg aatttaagcg ttggaccgcc gacgttgagg
5161 ctggtaagtg gctgcaagct aagggtgcta agccggctgc taaaccgcaa gaaacgagtg
5221 tcgctgatga ggctccggag gttaccgcta ccgttatctc ttacgataat tatgttacga
5281 ttctggacga ggaaacctta aaggcttgga tcgctaaatt agagaaggct cctgttttcg
5341 ctttcgacac ggaaacggat tctctggaca atattagtgc gaatcttgtt ggtctgagtt
5401 tcgcaattga accgggtgtt gctgcttaca tccctgtggc acacgactac ctggacgctc
5461 cggaccagat ttcacgtgaa cgcgctctgg aactgctgaa gcctttatta gaggacgaga
5521 aagctttgaa agttggtcag aatttgaagt atgctcgtgg aatcttagct aattatggta
5581 tcgagttgcg cggtatcgct ttcgacacga tgttggaatc ttatatcctg aactctgtcg
5641 ctggtcgcca tgacatggac tctctggctg agcgctggct gaaacataag acgattacct
5701 tcgaggaaat cgcaggaaag ggtaagaacc agctcacgtt caatcaaatc gctctggagg
5761 aagctggtcg ctatgctgct gaggacgctg acgttactct gcaactgcac ttgaagatgt
5821 ggcctgactt gcagaagcat aagggtccac tgaatgtttt tgaaaacatt gagatgcctt
5881 tggttccagt tctgtctcgt atcgagcgca atggcgttaa aattgaccca aaggttttac
5941 ataaccactc agaggaactg acgctgcgct tagccgaatt ggagaaaaag gctaccgaaa
6001 cgttcggctc gtggtatcag cctaaaggtg gcactgagat gttctgccat ccgcgaacag
6061 gtaagccact acctaaatac cctcgcatta agacacctaa agttggtggt atctttaaga
6121 agcctaagaa caaggcacag cgagaaggcc gtgagccttg cgaacttgat acccgcgagt
6181 acgttgctgg tgctccttac accccagttg aacatgttgt gttcaatctg tcatctacga
6241 aacaactgca gactatcctg ttcgagaagc aaggtatcaa gccattaaaa aagacccctg
6301 gcggtgctcc gtctacctct gaggaagttt tggaggagtt agctttggat taccctctgc
6361 cgaaggttat cttggaatac cgcggtttgg ctaaattgaa gtctacttat acggataaac
6421 ttcctttgat gattaatcca aagacgggtc gcgttcacac gtcgtaccat caagctgtta
6481 ccgctaccgg tcgcctgtct tctacggatc cgaatttaca gaatattcct gtgcgcaatg
6541 aggagggccg ccgcattcgt caagctttta tcgctccgga agactacgtt atcgtttctg
6601 ctgattattc tcaaaatgaa ttacgtatca tggctcacct gtctcgcgat aagggtctgt
6661 tgacggcctt tgctgagggt aaggacattc atcgtgctac cgctgctgag gttttcggcc
6721 tgccgttgga aacggttacg tctgaacagc gtcgctctgc taagcgtatt aatttcggct
6781 taatctacgg tatgtctgcg tttggcttag ctcgtcagct gaatatcccg cgcaaggaag
6841 ctcaaaaata tatggatctg tattttgagc gttacccggg tgttttggaa tacatggagc
6901 gtacgcgcgc gcaagctaag gaacaaggtt atgtggaaac cttggatggt cgtcgcttgt
6961 acttgcctga cattaagtct tctaacggcg cccgccgcgc tgctgccgag cgcgctgcta
7021 tcaatgctcc gatgcaaggt actgctgctg atattattaa gcgtgctatg atcgctgtgg
7081 acgcttggct gcaagctgaa cagcctcgcg ttcgcatgat tatgcaagtt catgacgagt
7141 tggttttcga ggtgcataag gacgacgtgg acgctgttgc taaacaaatc caccagttga
7201 tggagaattg cacgcgctta gacgttccgc tgctggttga agttggttct ggtgaaaact
7261 gggaccaggc tcactaataa tggcctcggt accaaagacg aacaataaga cgctgaaaag
7321 cgtctttttt cgttttggtc cgctgagcag ttacagagat gttacgaacc actagtgcac
7381 tgcagtacag tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
7441 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
7501 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
7561 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
7621 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
7681 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
7741 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgaggc gaaatacgcg
7801 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
7861 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
7921 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
7981 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
8041 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
8101 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
8161 atataaatca gcatccatgt tggaatttaa tcgcggcctg gagcaagacg tttcccgttg
8221 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
8281 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct
8341 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag gagttcatga ccaaaatccc
8401 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
8461 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
8521 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
8581 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
8641 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
8701 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
8761 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
8821 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
8881 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
8941 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
9001 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
9061 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
9121 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
9181 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
9241 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
9301 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
9361 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
9421 tgctcccggc atccgcttac agacaagctg tgaccttctc cgggagctgc atgtgtcaga
9481 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
9541 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
9601 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
9661 tggtcactga tgcctccgtg taagggggat ttctgttcat ccctgaattc gcatctagac
9721 tgaactggcc gataattgca gacgaacgga tttactttac agctagctca gtcctaggta
9781 ttatgctagc gctagcaaag aggagaaaag atctatggag aaaaaaatca ctggatatac
9841 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
9901 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
9961 gaaaaataag cacaagtttt aaccggcctt tattcacatt cttgcccgcc tgatgaatgc
10021 tcatccggaa tttcgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
10081 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata
10141 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
10201 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgttt cagccaatcc
10261 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
10321 cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
10381 tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca
10441 acagtactgc gatgagtggc agggcggggc gtaaggatcc taactcgagc tcggtaccaa
10501 attccagaaa agaggcctcc cgaaaggggg gccttttttc gttttggtcc gctgccaaac
10561 cagatgtcaa cacagctaca acgtttatag ctagctcagc ccttggtaca atgctagctg
10621 agacggaaag tgaaacgtga tttcatgcgt cattttgaac attttgtaaa tcttatttaa
10681 taatgtgtgc ggcaattcac atttaattta tgaatgtttt cttaacatcg cggcaactca
10741 agaaacggca ggttcggatc ttagctacta gagaaagagg agaaatacta gatgcgtaaa
10801 ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg tgatgtcaac
10861 ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg taaactgacg
10921 ctgaagttca tctgtactac tggtaaactg ccggttcctt ggccgactct ggtaacgacg
10981 ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca gcatgacttc
11041 ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt taaggatgac
11101 ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt aaaccgcatt
11161 gagctgaaag gcattgactt taaagaggac ggcaatatcc tgggccataa gctggaatac
11221 aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg cattaaagcg
11281 aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga tcactaccag
11341 caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta tctgagcacg
11401 caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct gctggagttc
11461 gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaatgacc aggcatcaaa
11521 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
11581 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatacgt
11641 ctcagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
11701 aagtggcacc gagtcggtgc tttttttgaa gcttgggccc gaacaaaaac tcatctcaga
11761 agaggatctg aatagcgccg tcgaccatca tcatcatcat cattgagttt aaacggactc
11821 cagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
11881 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgaggt ggtcccacct
11941 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggactccc
12001 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
12061 ggcctttcgt tttatctgtt gtttgtcggt gaacgacagg gctggatgca cacactggct
12121 taagatgaca acgttaagac ccactttcac atttaagttg tttttctaat ccgcatatga
12181 tcaattcaag gccgaataag aaggctggct ctgcaccttg gtgatcaaat aattcgatag
12241 cttgtcgtaa taatggcggc atactatcag tagtaggtgt ttccctttct tctttagcga
12301 cttgatgctc ttgatcttcc aatacgcaac ctaaagtaaa atgccccaca gcgctgagtg
12361 catataatgc attctctagt gaaaaacctt gttggcataa aaaggctaat tgattttcga
12421 gagtttcata ctgtttttct gtaggccgtg tacctaaatg tacttttgct ccatcgcgat
12481 gacttagtaa agcacatcta aaacttttag cgttattacg taaaaaatct tgccagcttt
12541 ccccttctaa agggcaaaag tgagtatggt gcctatctaa catctcaatg gctaaggcgt
12601 cgagcaaagc ccgcttattt tttacatgcc aatacaatgt aggctgctct acacctagct
12661 tctgggcgag tttacgggtt gttaaacctt cgattccgac ctcattaagc agctctaatg
12721 cgctgttaat cactttactt ttatctaatc tag
//
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