| 质粒名称: | pXPR_123 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 96919 |
| 存储实验室: | John Doench, David Root |
|
载体骨架基本信息 |
|
| 载体名称: | pXPR |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒 |
| 筛选标记: | 杀稻瘟菌素 |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | dCas9 |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | dCas9 |
| GenBank编号 | -- |
| 启动子 | EF1a |
| 标签/融合蛋白: | p300（插入件上的C端子） |
| 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: | pXPR_123 was a gift from John Doench & David Root (Addgene plasmid # 96919 ; http://n2t.net/addgene:96919 ; RRID:Addgene_96919) |
| 参考文献部分: | Orthologous CRISPR-Cas9 enzymes for combinatorial genetic screens. Najm FJ, Strand C, Donovan KF, Hegde M, Sanson KR, Vaimberg EW, Sullender ME, Hartenian E, Kalani Z, Fusi N, Listgarten J, Younger ST, Bernstein BE, Root DE, Doench JG. Nat Biotechnol. 2018 Feb;36(2):179-189. doi: 10.1038/nbt.4048. Epub 2017 Dec 18. 10.1038/nbt.4048 PubMed 29251726 |
买家导航
pXPR123载体基本信息
pXPR123载体图谱质粒图谱和多克隆位点信息
pXPR123载体简介
pXPR123载体序列
LOCUS pXPR_123 15807 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION for CRISPRi, lentiviral expression of dCas9-p300 2A BlastR..
ACCESSION .
VERSION .
KEYWORDS pXPR_123
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 15807)
AUTHORS Najm FJ, Strand C, Donovan KF, Hegde M, Sanson KR, Vaimberg EW,
Sullender ME, Hartenian E, Kalani Z, Fusi N, Listgarten J, Younger
ST, Bernstein BE, Root DE, Doench JG
TITLE Orthologous CRISPR-Cas9 enzymes for combinatorial genetic screens.
JOURNAL Nat Biotechnol. 2018 Feb;36(2):179-189. doi: 10.1038/nbt.4048. Epub
2017 Dec 18.
PUBMED 29251726
REFERENCE 2 (bases 1 to 15807)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..15807
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 248..676
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(335..354)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 545..566
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(685..705)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 690..1019
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 870..1005
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 932..951
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 1067..1114
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 1133..1507
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 1637..1758
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1674..1693)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1728..1747
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(1807..1823)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1807..1823)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1820..1842)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1831..1847
/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(1855..1885)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1900..1921
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2038..2055)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2209..2797)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2289..2308)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2968..3828)
/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 3591..3610
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3829..3933)
/gene="bla"
/label=AmpR promoter
primer_bind complement(4008..4027)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 4199..4578
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4580..4778
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 4733..4753
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 4796..4976
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 5023..5148
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5641..5874
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 6401..6518
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 6648..7826
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 6879..7817
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 7774..7794
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 7842..7889
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 7890..11990
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the N863A mutation in the HNH
catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 12045..12065
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 12081..13931
/codon_start=1
/gene="EP300"
/product="catalytic histone acetyltransferase core of the
human E1A-associated protein p300 (Delvecchio et al.,
2013)"
/label=p300 core
/translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI
VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ
EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG
ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG
FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA
SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP
PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY
IFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE
GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS
RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL
MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD"
regulatory 13695..13704
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 13932..13958
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 13983..14036
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 14037..14435
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
misc_feature 14460..15048
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(14513..14533)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(14931..14942)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(15051..15067)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(15052..15068)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 15573..15753
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(15779..15796)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 15785..202
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
ORIGIN
1 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
61 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
121 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
181 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
241 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
301 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
361 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
421 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
481 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
541 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
601 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
661 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
721 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
781 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
841 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
901 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
961 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
1021 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
1081 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
1141 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg
1201 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
1261 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
1321 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
1381 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
1441 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
1501 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
1561 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
1621 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
1681 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
1741 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
1801 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
1861 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
1921 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
1981 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
2041 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
2101 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
2161 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
2221 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
2281 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
2341 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
2401 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
2461 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
2521 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
2581 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
2641 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
2701 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
2761 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
2821 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
2881 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
2941 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
3001 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
3061 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
3121 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
3181 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
3241 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
3301 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
3361 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
3421 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
3481 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
3541 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
3601 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
3661 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
3721 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
3781 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
3841 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
3901 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
3961 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
4021 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
4081 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
4141 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
4201 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
4261 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
4321 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
4381 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
4441 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
4501 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
4561 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
4621 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
4681 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
4741 ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc
4801 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
4861 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
4921 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
4981 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
5041 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
5101 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
5161 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
5221 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
5281 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
5341 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
5401 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
5461 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
5521 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
5581 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
5641 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
5701 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
5761 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
5821 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
5881 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
5941 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
6001 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
6061 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
6121 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
6181 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
6241 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
6301 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg
6361 tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
6421 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
6481 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
6541 gagatccagt ttggttaatt agctagctgc aaagatggat aaagttttaa acagagagga
6601 atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc tccggtgccc
6661 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
6721 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
6781 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
6841 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
6901 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
6961 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
7021 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
7081 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
7141 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
7201 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
7261 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
7321 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
7381 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
7441 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
7501 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
7561 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
7621 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
7681 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
7741 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
7801 tttttcttcc atttcaggtg tcgtgacgta cggccaccat gaaaaggccg gcggccacga
7861 aaaaggccgg ccaggcaaaa aagaaaaagg acaagaagta cagcatcggc ctggccatcg
7921 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
7981 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
8041 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat
8101 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
8161 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
8221 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
8281 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
8341 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
8401 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
8461 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
8521 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
8581 agaagaagaa tggcctgttc ggcaacctga ttgccctgag cctgggcctg acccccaact
8641 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
8701 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
8761 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
8821 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
8881 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
8941 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
9001 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
9061 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
9121 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
9181 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
9241 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
9301 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
9361 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
9421 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
9481 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
9541 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
9601 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg
9661 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa
9721 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga
9781 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga
9841 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg
9901 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa
9961 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc
10021 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg
10081 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg
10141 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga
10201 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg
10261 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt
10321 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc
10381 ggctgtccga ctacgatgtg gaccacatcg tgcctcagag ctttctgaag gacgactcca
10441 tcgacaacaa ggtgctgacc agaagcgaca aggcccgggg caagagcgac aacgtgccct
10501 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga
10561 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg
10621 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg
10681 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg
10741 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt
10801 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg
10861 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg
10921 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg
10981 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
11041 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
11101 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
11161 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
11221 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
11281 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
11341 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
11401 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
11461 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
11521 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat
11581 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
11641 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
11701 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
11761 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
11821 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
11881 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
11941 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac agcgctggag
12001 gaggtggaag cggaggagga ggaagcggag gaggaggtag cggacctaag aaaaagagga
12061 aggtggcggc cgctggatcc attttcaaac cagaagaact acgacaggca ctgatgccaa
12121 ctttggaggc actttaccgt caggacccag aatcccttcc ctttcgtcaa cctgtggacc
12181 ctcagctttt aggaatccct gattactttg atattgtgaa gagccccatg gatctttcta
12241 ccattaagag gaagttagac actggacagt atcaggagcc ctggcagtat gtcgatgata
12301 tttggcttat gttcaataat gcctggttat ataaccggaa aacatcacgg gtatacaaat
12361 actgctccaa gctctctgag gtctttgaac aagaaattga cccagtgatg caaagccttg
12421 gatactgttg tggcagaaag ttggagttct ctccacagac actgtgttgc tacggcaaac
12481 agttgtgcac aatacctcgt gatgccactt attacagtta ccagaacagg tatcatttct
12541 gtgagaagtg tttcaatgag atccaagggg agagcgtttc tttgggggat gacccttccc
12601 agcctcaaac tacaataaat aaagaacaat tttccaagag aaaaaatgac acactggacc
12661 ctgaactgtt tgttgaatgt acggagtgcg gaagaaagat gcatcagatc tgtgtccttc
12721 accatgagat catctggcct gctggattcg tctgtgatgg ctgtttaaag aaaagtgcac
12781 gaactaggaa agaaaataag ttttctgcta aaaggttgcc atctaccaga cttggcacct
12841 ttctagagaa tcgtgtgaat gactttctga ggcgacagaa tcaccctgag tcaggagagg
12901 tcactgttag agtagttcat gcttctgaca aaaccgtgga agtaaaacca ggcatgaaag
12961 caaggtttgt ggacagtgga gagatggcag aatcctttcc ataccgaacc aaagccctct
13021 ttgcctttga agaaattgat ggtgttgacc tgtgcttctt tggcatgcat gttcaagagt
13081 atggctctga ctgccctcca cccaaccaga ggagagtata catatcttac ctcgatagtg
13141 ttcatttctt ccgtcctaaa tgcttgagga ctgcagtcta tcatgaaatc ctaattggat
13201 atttagaata tgtcaagaaa ttaggttaca caacagggca tatttgggca tgtccaccaa
13261 gtgagggaga tgattatatc ttccattgcc atcctcctga ccagaagata cccaagccca
13321 agcgactgca ggaatggtac aaaaaaatgc ttgacaaggc tgtatcagag cgtattgtcc
13381 atgactacaa ggatattttt aaacaagcta ctgaagatag attaacaagt gcaaaggaat
13441 tgccttattt cgagggtgat ttctggccca atgttctgga agaaagcatt aaggaactgg
13501 aacaggagga agaagagaga aaacgagagg aaaacaccag caatgaaagc acagatgtga
13561 ccaagggaga cagcaaaaat gctaaaaaga agaataataa gaaaaccagc aaaaataaga
13621 gcagcctgag taggggcaac aagaagaaac ccgggatgcc caatgtatct aacgacctct
13681 cacagaaact atatgccacc atggagaagc ataaagaggt cttctttgtg atccgcctca
13741 ttgctggccc tgctgccaac tccctgcctc ccattgttga tcctgatcct ctcatcccct
13801 gcgatctgat ggatggtcgg gatgcgtttc tcacgctggc aagggacaag cacctggagt
13861 tctcttcact ccgaagagcc cagtggtcca ccatgtgcat gctggtggag ctgcacacgc
13921 agagccagga ctacccgtac gacgttccgg actacgcttc tgatatctgt acaggcagtg
13981 gagagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct ggcccaatgg
14041 ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca
14101 gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca
14161 tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc
14221 tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg
14281 agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc
14341 ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg
14401 aattgctgcc ctctggttat gtgtgggagg gctaagaatt cgatatcaag cttatcggta
14461 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
14521 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
14581 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
14641 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
14701 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
14761 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
14821 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
14881 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
14941 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
15001 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
15061 gacctcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg
15121 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc
15181 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
15241 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt
15301 ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga
15361 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg
15421 tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga
15481 tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc
15541 acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag
15601 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
15661 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
15721 gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg ctgatcagcc
15781 tcgactgtgc cttctagttg ccagcca
//
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pXPR123载体质粒应用举例
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