| 质粒名称: | All_in_one_CRISPR/Cas9_LacZ |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 74293 |
| 存储实验室: | Lynne Postovit |
|
载体骨架基本信息 |
|
| 载体名称: | pCRISPR-CG01 |
| 载体出品公司: | Genecopedia |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 10415 bp |
| 完整质粒大小: | 10694 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，细菌表达，CRISP |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | LacZ-alpha |
| 物种来源: | Synthetic |
| 插入大小: | 279 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | Lac promoter |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gateway克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | SP6 Forward: ATTTAGGTGACACTATAGA |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | All_in_one_CRISPR/Cas9_LacZ was a gift from Lynne Postovit (Addgene plasmid # 74293 ; http://n2t.net/addgene:74293 ; RRID:Addgene_74293) |
| 参考文献部分: | A Digital PCR-Based Method for Efficient and Highly Specific Screening of Genome Edited Cells. Findlay SD, Vincent KM, Berman JR, Postovit LM. PLoS One. 2016 Apr 18;11(4):e0153901. doi: 10.1371/journal.pone.0153901. eCollection 2016. PONE-D-16-05005 [pii] PubMed 27089539 |
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 All_in_one_CRISPR/Cas9_LacZ 10694 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION ""All-in-one"" CRISPR/Cas9 (wt) plasmid for cloning of custom gRNA
with blue/white screening.
ACCESSION .
VERSION .
KEYWORDS All_in_one_CRISPR/Cas9_LacZ
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10694)
AUTHORS Findlay SD, Vincent KM, Berman JR, Postovit LM
TITLE A Digital PCR-Based Method for Efficient and Highly Specific
Screening of Genome Edited Cells.
JOURNAL PLoS One. 2016 Apr 18;11(4):e0153901. doi:
10.1371/journal.pone.0153901. eCollection 2016.
PUBMED 27089539
REFERENCE 2 (bases 1 to 10694)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10694
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 144..161
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 278..299
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 314..344
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 352..368
/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)."
primer_bind 376..392
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 412..430
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 412..429
/label=SP6
/note="SP6 promoter, forward primer"
promoter 442..460
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 591..611
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 762..781
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
protein_bind 825..846
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 861..891
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 899..915
/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)."
primer_bind 923..939
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 935..1213
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEW"
primer_bind 958..978
/label=T3
/note="T3 promoter, forward primer"
promoter 960..978
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter complement(1026..1044)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1054..1071)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1054..1070)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1063..1085)
/label=M13/pUC Forward
/note="In lacZ gene"
enhancer 1276..1655
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1656..1859
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1810..1830
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
promoter 1904..1922
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 1936..1945
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1942..6045
/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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 6061..6081
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6082..6108
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
primer_bind complement(6125..6145)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 6130..6459
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6310..6445
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6372..6391
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
regulatory 6517..6526
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 6523..7233
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(6668..6686)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 6936..6955
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 7249..7822
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(7416..7433)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
CDS 7858..8661
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(7921..7940)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 8531..8550
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 8793..8914
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8830..8849)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8884..8903
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(8963..8979)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8987..9003
/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 9065..9925
/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 complement(9283..9302)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 10096..10684
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10585..10604
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
61 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
121 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
181 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
241 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
301 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
361 gataacaatt tcacacagga aacagctatg acatgattac gaattgcaac gatttaggtg
421 acactataga agagaaggaa ttaatacgac tcactatagg gagagagaga gaattaccct
481 cactaaaggg aggagaagca tgaattcccc agtggaaaga cgcgcaggca aaacgcacca
541 cgtgacggag cgtgaccgcg cgccgagcgc gcgccaaggt cgggcaggaa gagggcctat
601 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattagaa
661 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
721 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
781 taactcgaga cgcggtggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
841 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
901 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
961 attaaccctc actaaaggga acaaaagctg gaggacctcg agggggggcc cggtacccaa
1021 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
1081 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
1141 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
1201 tggcgaatgg taggacgcgc cctgtagcgg cgcgtctcac gctgcttcgc gatgtacggg
1261 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
1321 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
1381 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
1441 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
1501 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
1561 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
1621 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
1681 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
1741 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
1801 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
1861 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
1921 gggagaccca agcttgccac catggacaag aagtacagca tcggcctgga catcggtacc
1981 aacagcgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaagttcaag
2041 gtgctgggca acaccgaccg ccacagcatc aagaagaacc tgatcggcgc cctgctgttc
2101 gacagcggcg agaccgccga ggccacccgc ctgaagcgca ccgcccgccg ccgctacacc
2161 cgccgcaaga accgcatctg ctacctgcag gagatcttca gcaacgagat ggccaaggtg
2221 gacgacagct tcttccaccg cctggaggag agcttcctgg tggaggagga caagaagcac
2281 gagcgccacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
2341 accatctacc acctgcgcaa gaagctggtg gacagcaccg acaaggccga cctgcgcctg
2401 atctacctgg ccctggccca catgatcaag ttccgcggcc acttcctgat cgagggcgac
2461 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
2521 cagctgttcg aggagaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgagc
2581 gcccgcctga gcaagagccg ccgcctggag aacctgatcg cccagctgcc cggcgagaag
2641 aagaacggcc tgttcggcaa cctgatcgcc ctgagcctgg gcctgacccc caacttcaag
2701 agcaacttcg acctggccga ggacgccaag ctgcagctga gcaaggacac ctacgacgac
2761 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt cctggccgcc
2821 aagaacctga gcgacgccat cctgctgagc gacatcctgc gcgtgaacac cgagatcacc
2881 aaggcccccc tgagcgccag catgatcaag cgctacgacg agcaccacca ggacctgacc
2941 ctgctgaagg ccctggtgcg ccagcagctg cccgagaagt acaaggagat cttcttcgac
3001 cagagcaaga acggctacgc cggctacatc gacggcggcg ccagccagga ggagttctac
3061 aagttcatca agcccatcct ggagaagatg gacggcaccg aggagctgct ggtgaagctg
3121 aaccgcgagg acctgctgcg caagcagcgc accttcgaca acggcagcat cccccaccag
3181 atccacctgg gcgagctgca cgccatcctg cgccgccagg aggacttcta ccccttcctg
3241 aaggacaacc gcgagaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
3301 cccctggccc gcggcaacag ccgcttcgcc tggatgaccc gcaagagcga ggagaccatc
3361 accccctgga acttcgagga ggtggtggac aagggcgcca gcgcccagag cttcatcgag
3421 cgcatgacca acttcgacaa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
3481 ctgtacgagt acttcaccgt gtacaacgag ctgaccaagg tgaagtacgt gaccgagggc
3541 atgcgcaagc ccgccttcct gagcggcgag cagaagaagg ccatcgtgga cctgctgttc
3601 aagaccaacc gcaaggtgac cgtgaagcag ctgaaggagg actacttcaa gaagatcgag
3661 tgcttcgaca gcgtggagat cagcggcgtg gaggaccgct tcaacgccag cctgggcacc
3721 taccacgacc tgctgaagat catcaaggac aaggacttcc tggacaacga ggagaacgag
3781 gacatcctgg aggacatcgt gctgaccctg accctgttcg aggaccgcga gatgatcgag
3841 gagcgcctga agacctacgc ccacctgttc gacgacaagg tgatgaagca gctgaagcgc
3901 cgccgctaca ccggctgggg ccgcctgagc cgcaagctta tcaacggcat ccgcgacaag
3961 cagagcggca agaccatcct ggacttcctg aagagcgacg gcttcgccaa ccgcaacttc
4021 atgcagctga tccacgacga cagcctgacc ttcaaggagg acatccagaa ggcccaggtg
4081 agcggccagg gcgacagcct gcacgagcac atcgccaacc tggccggcag ccccgccatc
4141 aagaagggca tcctgcagac cgtgaaggtg gtggacgagc tggtgaaggt gatgggccgc
4201 cacaagcccg agaacatcgt gatcgagatg gcccgcgaga accagaccac ccagaagggc
4261 cagaagaaca gccgcgagcg catgaagcgc atcgaggagg gcatcaagga gctgggcagc
4321 cagatcctga aggagcaccc cgtggagaac acccagctgc agaacgagaa gctgtacctg
4381 tactacctgc agaacggccg cgacatgtac gtggaccagg agctggacat caaccgcctg
4441 agcgactacg acgtggacca catcgtgccc cagagcttcc tgaaggacga cagcatcgac
4501 aacaaggtgc tgacccgcag cgacaagaac cgcggcaaga gcgacaacgt gcccagcgag
4561 gaggtggtga agaagatgaa gaactactgg cgccagctgc tgaacgccaa gctgatcacc
4621 cagcgcaagt tcgacaacct gaccaaggcc gagcgcggcg gcctgagcga gctggacaag
4681 gccggcttca tcaagcgcca gctggtggag acccgccaga tcaccaagca cgtggcccag
4741 atcctggaca gccgcatgaa caccaagtac gacgagaacg acaagctgat ccgcgaggtg
4801 aaggtgatca ccctgaagag caagctggtg agcgacttcc gcaaggactt ccagttctac
4861 aaggtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtggtg
4921 ggcaccgccc tgatcaagaa gtaccccaag ctggagagcg agttcgtgta cggcgactac
4981 aaggtgtacg acgtgcgcaa gatgatcgcc aagagcgagc aggagatcgg caaggccacc
5041 gccaagtact tcttctacag caacatcatg aacttcttca agaccgagat caccctggcc
5101 aacggcgaga tccgcaagcg ccccctgatc gagaccaacg gcgagaccgg cgagatcgtg
5161 tgggacaagg gccgcgactt cgccaccgtg cgcaaggtgc tgagcatgcc ccaggtgaac
5221 atcgtgaaga agaccgaggt gcagaccggc ggcttcagca aggagagcat cctgcccaag
5281 cgcaacagcg acaagctgat cgcccgcaag aaggactggg accccaagaa gtacggcggc
5341 ttcgacagcc ccaccgtggc ctacagcgtg ctggtggtgg ccaaggtgga gaagggcaag
5401 agcaagaagc tgaagagcgt gaaggagctg ctgggcatca ccatcatgga gcgcagcagc
5461 ttcgagaaga accccatcga cttcctggag gccaagggct acaaggaggt gaagaaggac
5521 ctgatcatca agctgcccaa gtacagcctg ttcgagctgg agaacggccg caagcgcatg
5581 ctggccagcg ccggcgagct gcagaagggc aacgagctgg ccctgcccag caagtacgtg
5641 aacttcctgt acctggccag ccactacgag aagctgaagg gcagccccga ggacaacgag
5701 cagaagcagc tgttcgtgga gcagcacaag cactacctgg acgagatcat cgagcagatc
5761 agcgagttca gcaagcgcgt gatcctggcc gacgccaacc tggacaaggt gctgagcgcc
5821 tacaacaagc accgcgacaa gcccatccgc gagcaggccg agaacatcat ccacctgttc
5881 accctgacca acctgggcgc ccccgccgcc ttcaagtact tcgacaccac catcgaccgc
5941 aagcgctaca ccagcaccaa ggaggtgctg gacgccaccc tgatccacca gagcatcacc
6001 ggtctgtacg agacccgcat cgacctgagc cagctgggcg gcgacggcgg ctccggacct
6061 ccaaagaaaa agagaaaagt atacccctac gacgtgcccg actacgccta ataactcgag
6121 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
6181 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
6241 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
6301 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
6361 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
6421 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
6481 tccattttcg gatctgatcg gcgcgccata ccggtcgcca ccatggtgag caagggcgag
6541 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
6601 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
6661 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
6721 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
6781 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
6841 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
6901 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
6961 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
7021 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
7081 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
7141 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
7201 tccaccggcg gcatggacga gctgtacaag taggcggccg cgatcccgcc cctctccctc
7261 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
7321 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
7381 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
7441 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
7501 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
7561 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
7621 gatagttgtg gaaagagtca aatggctcac ctcaagcgta ttcaacaagg ggctgaagga
7681 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
7741 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
7801 cctttgaaaa acacgatgat aagcttgcca caacccgcga taattcctgc agccaatatg
7861 ggatcggcca ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
7921 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
7981 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
8041 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
8101 gcagtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
8161 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
8221 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
8281 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
8341 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgtatg
8401 cccgacggcg atgatctcgt cgtgactcat ggcgatgcct gcttgccgaa tatcatggtg
8461 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
8521 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
8581 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
8641 cttcttgacg agttcttctg aacgcggtgc tacgagattt cgattccacc gccgccttct
8701 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
8761 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
8821 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
8881 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
8941 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
9001 caattccaca caacatacga gccggaagca taaaggtaag cctgaatatt gaaaaaggaa
9061 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
9121 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
9181 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
9241 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
9301 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
9361 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
9421 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
9481 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
9541 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
9601 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
9661 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
9721 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
9781 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
9841 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
9901 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
9961 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
10021 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
10081 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
10141 aaaaccaccg ctaccagcgg tggttttttt gccggatcaa gagctaccaa ctctttttcc
10201 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
10261 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
10321 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
10381 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
10441 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
10501 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
10561 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
10621 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
10681 gaaaaacgcc agca
//
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