| 质粒名称: | pCas12f1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 171182 |
| 存储实验室: | Kohsuke Honda |
|
载体骨架基本信息 |
|
| 载体名称: | pCas |
| 载体出品公司: | -- |
| 空载体大小: | 8576 bp |
| 完整质粒大小: | 12545 bp |
| 载体修饰: | 将靶向pTargetF的sgRNA序列替换为靶向pTarget12f1的序列。 |
| 载体类型: | 细菌表达,CRISPR |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素50 μg/mL |
| 生长温度: | 30 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Cas12f1,Cas14a1 |
| 物种来源: | Synthetic |
| 插入大小: | 1587 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | cas9 promoter from Streptococcus pyogenes |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | SalI/XhoI（克隆期间销毁） |
| 3’克隆位点: | KpnI(未破坏) |
| 5’测序引物 | GTATTGGGTAATATTTTTTGAAGAGATATTTTG |
| 3’测序引物 | CATTCAAATATGTATCCGCTCATG |
|
文献引用方法 |
|
| 材料和方法部分: | pCas12f1 was a gift from Kohsuke Honda (Addgene plasmid # 171182 ; http://n2t.net/addgene:171182 ; RRID:Addgene_171182) |
| 参考文献部分: | Genome editing by miniature CRISPR/Cas12f1 enzyme in Escherichia coli. Okano K, Sato Y, Hizume T, Honda K. J Biosci Bioeng. 2021 Aug;132(2):120-124. doi: 10.1016/j.jbiosc.2021.04.009. Epub 2021 May 19. 10.1016/j.jbiosc.2021.04.009 PubMed 34023220 |
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 pCas12f1 10163 bp ds-DNA circular SYN 29-JUN-2021
DEFINITION Constitutive expression of cas12f1, arabinose-inducible expression
of lambda recombinase, and IPTG-inducible expression of sgRNA for
curing of pTarget12f1.
ACCESSION .
VERSION .
KEYWORDS pCas12f1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10163)
TITLE Genome editing by miniature CRISPR/Cas12f1 enzyme in Escherichia
coli
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 10163)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 29, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10163
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 245..261
/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)."
CDS complement(530..1612)
/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 1574..1593
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(1613..1690)
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS complement(1823..2773)
/codon_start=1
/gene="rep101 encoding an A56V mutation"
/product="temperature-sensitive version of the RepA protein
needed for replication with the pSC101 origin (Armstrong et
al., 1984)"
/label=Rep101(Ts)
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 2821..3043
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
CDS complement(3695..4510)
/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 4418..4437
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 4502..4521
/label=pENTR-R
/note="pENTR vectors, reverse primer"
misc_RNA complement(4666..4744)
/label=tracrRNA
/note="trans-activating CRISPR RNA for the Streptococcus
pyogenes CRISPR/Cas9 system"
CDS complement(6771..7649)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRH
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 7676..7960
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 7883..7902
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
RBS 7987..7995
/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 8004..8420
/codon_start=1
/gene="gam"
/product="inhibitor of the host RecBCD nuclease in the
lambda Red system"
/label=Gam
/note="protects the double-stranded DNA substrate for
recombination"
/translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
CDS 8426..9211
/codon_start=1
/gene="bet"
/product="single-stranded DNA binding recombinase in the
lambda Red system"
/label=Beta
/note="mediates pairing of a single-stranded DNA with a
complementary target"
/translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
CDS 9208..9888
/codon_start=1
/gene="exo"
/product="5' to 3' double-stranded DNA exonuclease in the
lambda Red system"
/label=Exo
/note="creates 3' single-stranded DNA tails"
/translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
EQWR"
terminator 9889..10133
/label=-lambda tL3 terminator
/note="transcription terminator tL3 from phage lambda"
ORIGIN
1 gatctcaaaa aaacgccccc ctgacgagca tcagttgcat tccttcattc gtctattcgg
61 gttctgcaac tttcttgtgc agaattggag aggaaaaatg aatttgtttc gagggttact
121 ttccgaagaa agcacttctc gacattaggc tgatgcaagc agcccacctt cactcaagtt
181 ctaatcccct aagggacagc ttttggtgaa gcggttctcc actttatcag tgaagtgtgt
241 gaaattgtta tccgctcaca attccacaca acatacgagc cggatgatta attgtcaaca
301 gctcatttca gaatatttgc cagaaccgtt atgatgtcgg cgcaaaaaac attatccaga
361 acgggagtgc gccttgagcg acacgaatta tgcagtgatt tacgacctgc acagccatac
421 cacagcttcc gatggctgcc tgacgccaga agcattggtg caccgtgcag tcgatgataa
481 gctgtcaaac cagatcaatt cgcgctaact tacattaatt gcgttgcgct cactgcccgc
541 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
601 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
661 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
721 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
781 cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg
841 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
901 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
961 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
1021 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
1081 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
1141 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
1201 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
1261 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
1321 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
1381 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
1441 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
1501 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
1561 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
1621 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcaccatt
1681 cgatggtgtc aacgtaaatg catgccgctc gccttccatg ggtatggaca gttttccctt
1741 tgatatgtaa cggtgaacag ttgttctact tttgtttgtt agtcttgatg cttcactgat
1801 agatacaaga gccataagaa cctcagatcc ttccgtattt agccagtatg ttctctagtg
1861 tggttcgttg tttttgcgtg agccatgaga acgaaccatt gagatcatac ttactttgca
1921 tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt
1981 gtagtgtttt tcttagtccg ttatgtaggt aggaatctga tgtaatggtt gttggtattt
2041 tgtcaccatt catttttatc tggttgttct caagttcggt tacgagatcc atttgtctat
2101 ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca accaccaatt
2161 tcatattgct gtaagtgttt aaatctttac ttattggttt caaaacccat tggttaagcc
2221 ttttaaactc atggtagtta ttttcaagca ttaacatgaa cttaaattca tcaaggctaa
2281 tctctatatt tgccttgtga gttttctttt gtgttagttc ttttaataac cactcataaa
2341 tcctcataga gtatttgttt tcaaaagact taacatgttc cagattatat tttatgaatt
2401 tttttaactg gaaaagataa ggcaatatct cttcactaaa aactaattct aatttttcgc
2461 ttgagaactt ggcatagttt gtccactgga aaatctcaaa gcctttaacc aaaggattcc
2521 tgatttccac agttctcgtc atcagctctc tggttgcttt agctaataca ccataagcat
2581 tttccctact gatgttcatc atctgaacgt attggttata agtgaacgat accgtccgtt
2641 ctttccttgt agggttttca atcgtggggt tgagtagtgc cacacagcat aaaattagct
2701 tggtttcatg ctccgttaag tcatagcgac taatcgctag ttcatttgct ttgaaaacaa
2761 ctaattcaga catacatctc aattggtcta ggtgatttta atcactatac caattgagat
2821 gggctagtca atgataatta ctagtccttt tcctttgagt tgtgggtatc tgtaaattct
2881 gctagacctt tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat tccgctagac
2941 ctttgtgtgt tttttttgtt tatattcaag tggttataat ttatagaata aagaaagaat
3001 aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa ctcactactt tagtcagttc
3061 cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa
3121 accctaaagg cttaagtagc accctcgcaa gctcggttgc ggccgcaatc gggcaaatcg
3181 ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat
3241 tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgcct
3301 tttatggatt catgcaagga aactacccat aatacaagaa aagcccgtca cgggcttctc
3361 agggcgtttt atggcgggtc tgctatgtgg tgctatctga ctttttgctg ttcagcagtt
3421 cctgccctct gattttccag tctgaccact tcggattatc ccgtgacagg tcattcagac
3481 tggctaatgc acccagtaag gcagcggtat catcaacggg gtctgacgct cagtggaacg
3541 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
3601 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
3661 acagttacgt ttccaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat
3721 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
3781 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
3841 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
3901 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct
3961 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
4021 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat
4081 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
4141 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
4201 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
4261 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
4321 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
4381 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
4441 ataaatcagc atccatgttg gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa
4501 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
4561 atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt
4621 ccctgcaggg tttgcagtca gagtagaata gaagtatcaa aaaaagcacc gactcggtgc
4681 cactttttca agttgataac ggactagcct tattttaact tgctatgctg ttttgaatgg
4741 ttccaacaag attattttat aacttttata acaaataatc aaggagaaat tcaaagaaat
4801 ttatcagcca taaaacaata cttaatacta tagaatgata acaaaataaa ctacttttta
4861 aaagaatttt gtgttataat ctatttatta ttaagtattg ggtaatattt tttgaagaga
4921 tattttgaaa aagaaaaatt aaagcatatt aaactaattt cggaggtcat taaaactatt
4981 attgaaatca tcaaactcat tatggattta atttaaactt tttattttag gaggcaaaag
5041 tcgagatggc caaaaacacc attaccaaaa cactgaaact gcgtattgtg cgtccgtata
5101 atagcgcaga agtggaaaaa attgttgccg acgaaaaaaa caaccgcgaa aaaatcgcac
5161 tggaaaagaa caaagacaaa gtgaaagaag cctgcagcaa acatctgaaa gttgcagcat
5221 attgtaccac acaggttgaa cgtaatgcat gtctgttttg taaagcccgt aaactggatg
5281 acaagttcta tcaaaaactg cgtggtcagt ttccggatgc agttttttgg caagaaatca
5341 gcgaaatttt tcgccagctg cagaaacagg cagcagaaat ctataatcag agcctgatcg
5401 aactgtacta cgagattttt atcaaaggca aaggtattgc aaatgccagc agcgttgaac
5461 attatctgag tgatgtttgt tatacccgtg cagcagaact gtttaaaaac gcagcaattg
5521 caagcggtct gcgtagcaaa atcaaaagca attttcgtct gaaagaactg aaaaacatga
5581 aaagtggtct gccgaccacc aaaagcgata attttccgat tccgctggtt aaacagaaag
5641 gtggtcagta taccggtttt gaaattagca atcataatag cgacttcatc atcaagattc
5701 cgtttggtcg ttggcaggtc aaaaaagaga ttgataaata tcgtccgtgg gagaaatttg
5761 actttgaaca ggttcagaaa agcccgaaac cgattagcct gctgctgagc acccagcgtc
5821 gtaaacgtaa taaaggttgg agcaaagatg aaggcaccga agccgaaatc aaaaaagtta
5881 tgaatggcga ttatcagacc agctacattg aagttaaacg tggcagcaaa atcggtgaaa
5941 aaagcgcatg gatgctgaat ctgagcattg atgttccgaa aattgataaa ggtgtggatc
6001 cgagcattat tggtggtatt gatgttggtg ttaaatcacc gctggtttgc gcaattaaca
6061 atgcatttag ccgttatagc atcagcgata acgacctgtt tcacttcaac aagaaaatgt
6121 ttgcacgtcg tcgtatcctg ctgaaaaaaa accgtcataa acgtgcaggt catggtgcaa
6181 aaaacaaact gaaaccgatc accattctga ccgaaaaaag tgaacgtttt cgcaaaaagc
6241 tgattgaacg ttgggcatgt gaaatcgcgg atttcttcat taaaaacaaa gttggcaccg
6301 tgcagatgga aaatctggaa agcatgaaac gtaaagagga cagctatttt aacattcgcc
6361 tgcgtggctt ttggccgtat gcagaaatgc agaacaaaat cgagtttaag ctgaaacagt
6421 atggcatcga aattcgtaaa gtggcaccga ataataccag caaaacctgt agcaaatgtg
6481 gccatctgaa caactatttc aacttcgagt accgcaagaa aaacaaattc ccgcacttta
6541 aatgcgaaaa atgcaacttc aaagaaaacg ccgattataa tgcagccctg aatatttcaa
6601 acccgaaact gaaaagcacc aaagaggaac cgtaaggtac cagtatattt tagatgaaga
6661 ttatttctta atctagacat gagcggatac atatttgaat gtatttagaa aaataaacaa
6721 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg catcgattta ttatgacaac
6781 ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc
6841 ccggtgcatt ttttaaatac ccgcgagaag tagagttgat cgtcaaaacc aacattgcga
6901 ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt
6961 tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga
7021 cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc
7081 aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc
7141 gactcgttaa tcgcttccat gtgccgcagt aacaattgct caagcagatt tatcgccagc
7201 agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg
7261 aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc
7321 cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat
7381 tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata
7441 tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg
7501 gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa
7561 ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc
7621 agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt cagagaagaa
7681 accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg
7741 ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc
7801 atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat
7861 ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt agcggatcct
7921 acctgacgct ttttatcgca actctctact gtttctccat acccgttttt ttgggaattc
7981 gagctctaag gaggttataa aaaatggata ttaatactga aactgagatc aagcaaaagc
8041 attcactaac cccctttcct gttttcctaa tcagcccggc atttcgcggg cgatattttc
8101 acagctattt caggagttca gccatgaacg cttattacat tcaggatcgt cttgaggctc
8161 agagctgggc gcgtcactac cagcagctcg cccgtgaaga gaaagaggca gaactggcag
8221 acgacatgga aaaaggcctg ccccagcacc tgtttgaatc gctatgcatc gatcatttgc
8281 aacgccacgg ggccagcaaa aaatccatta cccgtgcgtt tgatgacgat gttgagtttc
8341 aggagcgcat ggcagaacac atccggtaca tggttgaaac cattgctcac caccaggttg
8401 atattgattc agaggtataa aacgaatgag tactgcactc gcaacgctgg ctgggaagct
8461 ggctgaacgt gtcggcatgg attctgtcga cccacaggaa ctgatcacca ctcttcgcca
8521 gacggcattt aaaggtgatg ccagcgatgc gcagttcatc gcattactga tcgttgccaa
8581 ccagtacggc cttaatccgt ggacgaaaga aatttacgcc tttcctgata agcagaatgg
8641 catcgttccg gtggtgggcg ttgatggctg gtcccgcatc atcaatgaaa accagcagtt
8701 tgatggcatg gactttgagc aggacaatga atcctgtaca tgccggattt accgcaagga
8761 ccgtaatcat ccgatctgcg ttaccgaatg gatggatgaa tgccgccgcg aaccattcaa
8821 aactcgcgaa ggcagagaaa tcacggggcc gtggcagtcg catcccaaac ggatgttacg
8881 tcataaagcc atgattcagt gtgcccgtct ggccttcgga tttgctggta tctatgacaa
8941 ggatgaagcc gagcgcattg tcgaaaatac tgcatacact gcagaacgtc agccggaacg
9001 cgacatcact ccggttaacg atgaaaccat gcaggagatt aacactctgc tgatcgccct
9061 ggataaaaca tgggatgacg acttattgcc gctctgttcc cagatatttc gccgcgacat
9121 tcgtgcatcg tcagaactga cacaggccga agcagtaaaa gctcttggat tcctgaaaca
9181 gaaagccgca gagcagaagg tggcagcatg acaccggaca ttatcctgca gcgtaccggg
9241 atcgatgtga gagctgtcga acagggggat gatgcgtggc acaaattacg gctcggcgtc
9301 atcaccgctt cagaagttca caacgtgata gcaaaacccc gctccggaaa gaagtggcct
9361 gacatgaaaa tgtcctactt ccacaccctg cttgctgagg tttgcaccgg tgtggctccg
9421 gaagttaacg ctaaagcact ggcctgggga aaacagtacg agaacgacgc cagaaccctg
9481 tttgaattca cttccggcgt gaatgttact gaatccccga tcatctatcg cgacgaaagt
9541 atgcgtaccg cctgctctcc cgatggttta tgcagtgacg gcaacggcct tgaactgaaa
9601 tgcccgttta cctcccggga tttcatgaag ttccggctcg gtggtttcga ggccataaag
9661 tcagcttaca tggcccaggt gcagtacagc atgtgggtga cgcgaaaaaa tgcctggtac
9721 tttgccaact atgacccgcg tatgaagcgt gaaggcctgc attatgtcgt gattgagcgg
9781 gatgaaaagt acatggcgag ttttgacgag atcgtgccgg agttcatcga aaaaatggac
9841 gaggcactgg ctgaaattgg ttttgtattt ggggagcaat ggcgatgacg catcctcacg
9901 ataatatccg ggtaggcgca atcactttcg tctactccgt tacaaagcga ggctgggtat
9961 ttcccggcct ttctgttatc cgaaatccac tgaaagcaca gcggctggct gaggagataa
10021 ataataaacg aggggctgta tgcacaaagc atcttctgtt gagttaagaa cgagtatcga
10081 gatggcacat agccttgctc aaattggaat caggtttgtg ccaataccag tagaaacaga
10141 cgaagaatcc atgggtatgg aca
//
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