| 质粒名称: | NL4-3 mCherry Luciferase |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 44965 |
| 存储实验室: | Warner Greene |
|
载体骨架基本信息 |
|
| 载体名称: | pNLENG1 |
| 载体出品公司: | AIDS Research and Reference Reagent Program |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 12424 bp |
| 载体修饰: | -- |
| 载体类型: | 慢病毒载体 |
| 筛选标记: | mCherry, Luciferase |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | mCherry, Luciferase, IRES,HIV 1 reporter virus |
| 物种来源: | Lentivirus, Human specific |
| 插入大小: | -- |
| 基因突变: | S34C mutation in Nef |
| GenBank编号 | -- |
| 启动子 | EF-1 alpha– driven mCherry:Puromycin |
| 标签/融合蛋白: |
带D212G的mCherry（插接件上的C端子）
荧光素酶，IRES（C端子插入）
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI (未破坏) |
| 3' 克隆位点: | XhoI（在克隆过程中被破坏） |
| 5' 测序引物 | from pNLENG1 5' |
| 3' 测序引物 | from pNLENG1 3' |
|
文献引用方法 |
|
| 材料和方法部分: | NL4-3 mCherry Luciferase was a gift from Warner Greene (Addgene plasmid # 44965 ; http://n2t.net/addgene:44965 ; RRID:Addgene_44965) |
| 参考文献部分: | A flexible model of HIV-1 latency permitting evaluation of many primary CD4 T-cell reservoirs. Lassen KG, Hebbeler AM, Bhattacharyya D, Lobritz MA, Greene WC. PLoS One. 2012;7(1):e30176. doi: 10.1371/journal.pone.0030176. Epub 2012 Jan 24. 10.1371/journal.pone.0030176 PubMed 22291913 |
买家导航
NL4-3 mCherry Luciferase载体质粒基本信息
NL4-3 mCherry Luciferase质粒图谱载体图谱和NL4-3 mCherry Luciferase载体序列质粒序列多克隆位点信息
NL4-3 mCherry Luciferase质粒载体简介
NL4-3 mCherry Luciferase质粒序列载体序列
LOCUS NL4-3 mCherry Luciferase 17833 bp ds-DNA circular SYN 16-DEC-2019
DEFINITION HIV dual reporter vector expressing mCh.
ACCESSION .
VERSION .
KEYWORDS NL4-3 mCherry Luciferase
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 17833)
AUTHORS Lassen KG, Hebbeler AM, Bhattacharyya D, Lobritz MA, Greene WC
TITLE A flexible model of HIV-1 latency permitting evaluation of many
primary CD4 T-cell reservoirs.
JOURNAL PLoS One. 2012;7(1):e30176. doi: 10.1371/journal.pone.0030176. Epub
2012 Jan 24.
PUBMED 22291913
REFERENCE 2 (bases 1 to 17833)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 16, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..17833
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 377..634
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
CDS 790..2292
/codon_start=1
/gene="gag"
/product="gag protein from human immunodeficiency virus 1"
/label=HIV-1 gag
/translation="MGARASVLSGGELDKWEKIRLRPGGKKQYKLKHIVWASRELERFA
VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTIAVLYCVHQRIDVKDTKEALDKI
EEEQNKSKKKAQQAAADTGNNSQVSQNYPIVQNLQGQMVHQAISPRTLNAWVKVVEEKA
FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRLHPVHAGP
IAPGQMREPRGSDIAGTTSTLQEQIGWMTHNPPIPVGEIYKRWIILGLNKIVRMYSPTS
ILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPG
ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNPATIMIQKGNFRNQRKTVKCFNCGKEG
HIAKNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSHKGRPGNFLQSRPEPTAP
PEESFRFGEETTTPSQKQEPIDKELYPLASLRSLFGSDPSSQ"
CDS 2085..5096
/codon_start=1
/gene="pol"
/product="pol protein from human immunodeficiency virus 1"
/label=HIV-1 pol
/translation="FFREDLAFPQGKAREFSSEQTRANSPTRRELQVWGRDNNSLSEAG
ADRQGTVSFSFPQITLWQRPLVTIKIGGQLKEALLDTGADDTVLEEMNLPGRWKPKMIG
GIGGFIKVRQYDQILIEICGHKAIGTVLVGPTPVNIIGRNLLTQIGCTLNFPISPIETV
PVKLKPGMDGPKVKQWPLTEEKIKALVEICTEMEKEGKISKIGPENPYNTPVFAIKKKD
STKWRKLVDFRELNKRTQDFWEVQLGIPHPAGLKQKKSVTVLDVGDAYFSVPLDKDFRK
YTAFTIPSINNETPGIRYQYNVLPQGWKGSPAIFQCSMTKILEPFRKQNPDIVIYQYMD
DLYVGSDLEIGQHRTKIEELRQHLLRWGFTTPDKKHQKEPPFLWMGYELHPDKWTVQPI
VLPEKDSWTVNDIQKLVGKLNWASQIYAGIKVRQLCKLLRGTKALTEVVPLTEEAELEL
AENREILKEPVHGVYYDPSKDLIAEIQKQGQGQWTYQIYQEPFKNLKTGKYARMKGAHT
NDVKQLTEAVQKIATESIVIWGKTPKFKLPIQKETWEAWWTEYWQATWIPEWEFVNTPP
LVKLWYQLEKEPIIGAETFYVDGAANRETKLGKAGYVTDRGRQKVVPLTDTTNQKTELQ
AIHLALQDSGLEVNIVTDSQYALGIIQAQPDKSESELVSQIIEQLIKKEKVYLAWVPAH
KGIGGNEQVDKLVSAGIRKVLFLDGIDKAQEEHEKYHSNWRAMASDFNLPPVVAKEIVA
SCDKCQLKGEAMHGQVDCSPGIWQLDCTHLEGKVILVAVHVASGYIEAEVIPAETGQET
AYFLLKLAGRWPVKTVHTDNGSNFTSTTVKAACWWAGIKQEFGIPYNPQSQGVIESMNK
ELKKIIGQVRDQAEHLKTAVQMAVFIHNFKRKGGIGGYSAGERIVDIIATDIQTKELQK
QITKIQNFRVYYRDSRDPVWKGPAKLLWKGEGAVVIQDNSDIKVVPRRKAKIIRDYGKQ
MAGDDCVASRQDED"
misc_feature 4781..4898
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
misc_feature 7759..7992
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 8515..8541
/codon_start=1
/product="nuclear export signal from the HIV Rev protein
(Fischer et al., 1995)"
/label=NES
/translation="LPPLERLTL"
CDS 8787..9494
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEGYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(8932..8950)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 9200..9219
/label=mCherry-F
/note="mCherry, forward primer"
CDS 9501..9554
/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 9558..11207
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind complement(9621..9639)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
misc_feature 11212..11794
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(11378..11395)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 11605..11624
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
LTR 12084..12717
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(14357..14374)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(14528..15116)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(14608..14627)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(15287..16147)
/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 15910..15929
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(16148..16252)
/gene="bla"
/label=AmpR promoter
primer_bind 16320..16338
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(16376..16398)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 16498..16517
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 tggaagggct aatttggtcc caaaaaagac aagagatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc agatatccac
121 tgacctttgg atggtgcttc aagttagtac cagttgaacc agagcaagta gaagaggcca
181 atgaaggaga gaacaacagc ttgttacacc ctatgagcca gcatgggatg gaggacccgg
241 agggagaagt attagtgtgg aagtttgaca gcctcctagc atttcgtcac atggcccgag
301 agctgcatcc ggagtactac aaagactgct gacatcgagc tttctacaag ggactttccg
361 ctggggactt tccagggagg tgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 gctacatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgctca aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagtaaa gccagaggag atctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcggta ttaagcgggg gagaattaga taaatgggaa
841 aaaattcggt taaggccagg gggaaagaaa caatataaac taaaacatat agtatgggca
901 agcagggagc tagaacgatt cgcagttaat cctggccttt tagagacatc agaaggctgt
961 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
1021 ttatataata caatagcagt cctctattgt gtgcatcaaa ggatagatgt aaaagacacc
1081 aaggaagcct tagataagat agaggaagag caaaacaaaa gtaagaaaaa ggcacagcaa
1141 gcagcagctg acacaggaaa caacagccag gtcagccaaa attaccctat agtgcagaac
1201 ctccaggggc aaatggtaca tcaggccata tcacctagaa ctttaaatgc atgggtaaaa
1261 gtagtagaag agaaggcttt cagcccagaa gtaataccca tgttttcagc attatcagaa
1321 ggagccaccc cacaagattt aaataccatg ctaaacacag tggggggaca tcaagcagcc
1381 atgcaaatgt taaaagagac catcaatgag gaagctgcag aatgggatag attgcatcca
1441 gtgcatgcag ggcctattgc accaggccag atgagagaac caaggggaag tgacatagca
1501 ggaactacta gtacccttca ggaacaaata ggatggatga cacataatcc acctatccca
1561 gtaggagaaa tctataaaag atggataatc ctgggattaa ataaaatagt aagaatgtat
1621 agccctacca gcattctgga cataagacaa ggaccaaagg aaccctttag agactatgta
1681 gaccgattct ataaaactct aagagccgag caagcttcac aagaggtaaa aaattggatg
1741 acagaaacct tgttggtcca aaatgcgaac ccagattgta agactatttt aaaagcattg
1801 ggaccaggag cgacactaga agaaatgatg acagcatgtc agggagtggg gggacccggc
1861 cataaagcaa gagttttggc tgaagcaatg agccaagtaa caaatccagc taccataatg
1921 atacagaaag gcaattttag gaaccaaaga aagactgtta agtgtttcaa ttgtggcaaa
1981 gaagggcaca tagccaaaaa ttgcagggcc cctaggaaaa agggctgttg gaaatgtgga
2041 aaggaaggac accaaatgaa agattgtact gagagacagg ctaatttttt agggaagatc
2101 tggccttccc acaagggaag gccagggaat tttcttcaga gcagaccaga gccaacagcc
2161 ccaccagaag agagcttcag gtttggggaa gagacaacaa ctccctctca gaagcaggag
2221 ccgatagaca aggaactgta tcctttagct tccctcagat cactctttgg cagcgacccc
2281 tcgtcacaat aaagataggg gggcaattaa aggaagctct attagataca ggagcagatg
2341 atacagtatt agaagaaatg aatttgccag gaagatggaa accaaaaatg atagggggaa
2401 ttggaggttt tatcaaagta agacagtatg atcagatact catagaaatc tgcggacata
2461 aagctatagg tacagtatta gtaggaccta cacctgtcaa cataattgga agaaatctgt
2521 tgactcagat tggctgcact ttaaattttc ccattagtcc tattgagact gtaccagtaa
2581 aattaaagcc aggaatggat ggcccaaaag ttaaacaatg gccattgaca gaagaaaaaa
2641 taaaagcatt agtagaaatt tgtacagaaa tggaaaagga aggaaaaatt tcaaaaattg
2701 ggcctgaaaa tccatacaat actccagtat ttgccataaa gaaaaaagac agtactaaat
2761 ggagaaaatt agtagatttc agagaactta ataagagaac tcaagatttc tgggaagttc
2821 aattaggaat accacatcct gcagggttaa aacagaaaaa atcagtaaca gtactggatg
2881 tgggcgatgc atatttttca gttcccttag ataaagactt caggaagtat actgcattta
2941 ccatacctag tataaacaat gagacaccag ggattagata tcagtacaat gtgcttccac
3001 agggatggaa aggatcacca gcaatattcc agtgtagcat gacaaaaatc ttagagcctt
3061 ttagaaaaca aaatccagac atagtcatct atcaatacat ggatgatttg tatgtaggat
3121 ctgacttaga aatagggcag catagaacaa aaatagagga actgagacaa catctgttga
3181 ggtggggatt taccacacca gacaaaaaac atcagaaaga acctccattc ctttggatgg
3241 gttatgaact ccatcctgat aaatggacag tacagcctat agtgctgcca gaaaaggaca
3301 gctggactgt caatgacata cagaaattag tgggaaaatt gaattgggca agtcagattt
3361 atgcagggat taaagtaagg caattatgta aacttcttag gggaaccaaa gcactaacag
3421 aagtagtacc actaacagaa gaagcagagc tagaactggc agaaaacagg gagattctaa
3481 aagaaccggt acatggagtg tattatgacc catcaaaaga cttaatagca gaaatacaga
3541 agcaggggca aggccaatgg acatatcaaa tttatcaaga gccatttaaa aatctgaaaa
3601 caggaaagta tgcaagaatg aagggtgccc acactaatga tgtgaaacaa ttaacagagg
3661 cagtacaaaa aatagccaca gaaagcatag taatatgggg aaagactcct aaatttaaat
3721 tacccataca aaaggaaaca tgggaagcat ggtggacaga gtattggcaa gccacctgga
3781 ttcctgagtg ggagtttgtc aatacccctc ccttagtgaa gttatggtac cagttagaga
3841 aagaacccat aataggagca gaaactttct atgtagatgg ggcagccaat agggaaacta
3901 aattaggaaa agcaggatat gtaactgaca gaggaagaca aaaagttgtc cccctaacgg
3961 acacaacaaa tcagaagact gagttacaag caattcatct agctttgcag gattcgggat
4021 tagaagtaaa catagtgaca gactcacaat atgcattggg aatcattcaa gcacaaccag
4081 ataagagtga atcagagtta gtcagtcaaa taatagagca gttaataaaa aaggaaaaag
4141 tctacctggc atgggtacca gcacacaaag gaattggagg aaatgaacaa gtagataaat
4201 tggtcagtgc tggaatcagg aaagtactat ttttagatgg aatagataag gcccaagaag
4261 aacatgagaa atatcacagt aattggagag caatggctag tgattttaac ctaccacctg
4321 tagtagcaaa agaaatagta gccagctgtg ataaatgtca gctaaaaggg gaagccatgc
4381 atggacaagt agactgtagc ccaggaatat ggcagctaga ttgtacacat ttagaaggaa
4441 aagttatctt ggtagcagtt catgtagcca gtggatatat agaagcagaa gtaattccag
4501 cagagacagg gcaagaaaca gcatacttcc tcttaaaatt agcaggaaga tggccagtaa
4561 aaacagtaca tacagacaat ggcagcaatt tcaccagtac tacagttaag gccgcctgtt
4621 ggtgggcggg gatcaagcag gaatttggca ttccctacaa tccccaaagt caaggagtaa
4681 tagaatctat gaataaagaa ttaaagaaaa ttataggaca ggtaagagat caggctgaac
4741 atcttaagac agcagtacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
4801 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
4861 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
4921 gagatccagt ttggaaagga ccagcaaagc tcctctggaa aggtgaaggg gcagtagtaa
4981 tacaagataa tagtgacata aaagtagtgc caagaagaaa agcaaagatc atcagggatt
5041 atggaaaaca gatggcaggt gatgattgtg tggcaagtag acaggatgag gattaacaca
5101 tggaaaagat tagtaaaaca ccatatgtat atttcaagga aagctaagga ctggttttat
5161 agacatcact atgaaagtac taatccaaaa ataagttcag aagtacacat cccactaggg
5221 gatgctaaat tagtaataac aacatattgg ggtctgcata caggagaaag agactggcat
5281 ttgggtcagg gagtctccat agaatggagg aaaaagagat atagcacaca agtagaccct
5341 gacctagcag accaactaat tcatctgcac tattttgatt gtttttcaga atctgctata
5401 agaaatacca tattaggacg tatagttagt cctaggtgtg aatatcaagc aggacataac
5461 aaggtaggat ctctacagta cttggcacta gcagcattaa taaaaccaaa acagataaag
5521 ccacctttgc ctagtgttag gaaactgaca gaggacagat ggaacaagcc ccagaagacc
5581 aagggccaca gagggagcca tacaatgaat ggacactaga gcttttagag gaacttaaga
5641 gtgaagctgt tagacatttt cctaggatat ggctccataa cttaggacaa catatctatg
5701 aaacttacgg ggatacttgg gcaggagtgg aagccataat aagaattctg caacaactgc
5761 tgtttatcca tttcagaatt gggtgtcgac atagcagaat aggcgttact cgacagagga
5821 gagcaagaaa tggagccagt agatcctaga ctagagccct ggaagcatcc aggaagtcag
5881 cctaaaactg cttgtaccaa ttgctattgt aaaaagtgtt gctttcattg ccaagtttgt
5941 ttcatgacaa aagccttagg catctcctat ggcaggaaga agcggagaca gcgacgaaga
6001 gctcatcaga acagtcagac tcatcaagct tctctatcaa agcagtaagt agtacatgta
6061 atgcaaccta taatagtagc aatagtagca ttagtagtag caataataat agcaatagtt
6121 gtgtggtcca tagtaatcat agaatatagg aaaatattaa gacaaagaaa aatagacagg
6181 ttaattgata gactaataga aagagcagaa gacagtggca atgagagtga aggagaagta
6241 tcagcacttg tggagatggg ggtggaaatg gggcaccatg ctccttggga tattgatgat
6301 ctgtagtgct acagaaaaat tgtgggtcac agtctattat ggggtacctg tgtggaagga
6361 agcaaccacc actctatttt gtgcatcaga tgctaaagca tatgatacag aggtacataa
6421 tgtttgggcc acacatgcct gtgtacccac agaccccaac ccacaagaag tagtattggt
6481 aaatgtgaca gaaaatttta acatgtggaa aaatgacatg gtagaacaga tgcatgagga
6541 tataatcagt ttatgggatc aaagcctaaa gccatgtgta aaattaaccc cactctgtgt
6601 tagtttaaag tgcactgatt tgaagaatga tactaatacc aatagtagta gcgggagaat
6661 gataatggag aaaggagaga taaaaaactg ctctttcaat atcagcacaa gcataagaga
6721 taaggtgcag aaagaatatg cattctttta taaacttgat atagtaccaa tagataatac
6781 cagctatagg ttgataagtt gtaacacctc agtcattaca caggcctgtc caaaggtatc
6841 ctttgagcca attcccatac attattgtgc cccggctggt tttgcgattc taaaatgtaa
6901 taataagacg ttcaatggaa caggaccatg tacaaatgtc agcacagtac aatgtacaca
6961 tggaatcagg ccagtagtat caactcaact gctgttaaat ggcagtctag cagaagaaga
7021 tgtagtaatt agatctgcca atttcacaga caatgctaaa accataatag tacagctgaa
7081 cacatctgta gaaattaatt gtacaagacc caacaacaat acaagaaaaa gtatccgtat
7141 ccagagggga ccagggagag catttgttac aataggaaaa ataggaaata tgagacaagc
7201 acattgtaac attagtagag caaaatggaa tgccacttta aaacagatag ctagcaaatt
7261 aagagaacaa tttggaaata ataaaacaat aatctttaag caatcctcag gaggggaccc
7321 agaaattgta acgcacagtt ttaattgtgg aggggaattt ttctactgta attcaacaca
7381 actgtttaat agtacttggt ttaatagtac ttggagtact gaagggtcaa ataacactga
7441 aggaagtgac acaatcacac tcccatgcag aataaaacaa tttataaaca tgtggcagga
7501 agtaggaaaa gcaatgtatg cccctcccat cagtggacaa attagatgtt catcaaatat
7561 tactgggctg ctattaacaa gagatggtgg taataacaac aatgggtccg agatcttcag
7621 acctggagga ggcgatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
7681 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
7741 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
7801 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctgatatagt
7861 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
7921 agtctggggc atcaaacagc tccaggcaag aatcctggct gtggaaagat acctaaagga
7981 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
8041 ttggaatgct agttggagta ataaatctct ggaacagatt tggaataaca tgacctggat
8101 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
8161 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
8221 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
8281 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
8341 gcagggatat tcaccattat cgtttcagac ccacctccca atcccgaggg gacccgacag
8401 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
8461 gaacggatcc ttagcactta tctgggacga tctgcggagc ctgtgcctct tcagctacca
8521 ccgcttgaga gacttactct tgattgtaac gaggattgtg gaacttctgg gacgcagggg
8581 gtgggaagcc ctcaaatatt ggtggaatct cctacagtat tggagtcagg aactaaagaa
8641 tagtgctgtt aacttgctca atgccacagc catagcagta gctgagggga cagatagggt
8701 tatagaagta ttacaagcag cttatagagc tattcgccac atacctagaa gaataagaca
8761 gggcttggaa aggattttgc tataagatgg tgagcaaggg cgaggaggat aacatggcca
8821 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
8881 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
8941 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
9001 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
9061 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
9121 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
9181 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
9241 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
9301 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
9361 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
9421 gctacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
9481 acgagctgta caagccccgg gagggcagag gaagtcttct aacatgcggt gacgtggagg
9541 agaatcccgg ccctcgagaa gacgccaaaa acataaagaa aggcccggcg ccattctatc
9601 cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg
9661 ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact tacgctgagt
9721 acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc
9781 acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt
9841 tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca
9901 acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa
9961 ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg gattctaaaa
10021 cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt
10081 ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt gcactgatca
10141 tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat agaactgcct
10201 gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg
10261 cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt
10321 tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg tttctgagga
10381 gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc tccttcttcg
10441 ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctggtg
10501 gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat ctgccaggta
10561 tcaggcaagg atatgggctc actgagacta catcagctat tctgattacg cccgaggggg
10621 atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc
10681 tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg agaggtccta
10741 tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg
10801 gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc ttcatcgttg
10861 accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct gaattggaat
10921 ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt cccgacgatg
10981 acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa
11041 aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag
11101 ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca
11161 gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgtaagga gcccctctcc
11221 ctcccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
11281 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
11341 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
11401 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
11461 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
11521 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
11581 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
11641 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
11701 acatgtgttt agtcgaggtt aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
11761 tcctttgaaa aacacgatga taatggccac aaccatgggt ggcaagtggt caaaaagtag
11821 tgtgattgga tggcctgctg taagggaaag aatgagacga gctgagccag cagcagatgg
11881 ggtgggagca gtatgtcgag acctagaaaa acatggagca atcacaagta gcaatacagc
11941 agctaacaat gctgcttgtg cctggctaga agcacaagag gaggaagagg tgggttttcc
12001 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
12061 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaaagaa gacaagatat
12121 ccttgatctg tggatctacc acacacaagg ctacttccct gattggcaga actacacacc
12181 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
12241 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag
12301 cctgcatgga atggatgacc ctgagagaga agtgttagag tggaggtttg acagccgcct
12361 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgacatcg
12421 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac
12481 tggggagtgg cgagccctca gatgctgcat ataagcagct gctttttgcc tgtactgggt
12541 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
12601 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
12661 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaccc
12721 aggaggtaga ggttgcagtg agccaagatc gcgccactgc attccagcct gggcaagaaa
12781 acaagactgt ctaaaataat aataataagt taagggtatt aaatatattt atacatggag
12841 gtcataaaaa tatatatatt tgggctgggc gcagtggctc acacctgcgc ccggcccttt
12901 gggaggccga ggcaggtgga tcacctgagt ttgggagttc cagaccagcc tgaccaacat
12961 ggagaaaccc cttctctgtg tatttttagt agattttatt ttatgtgtat tttattcaca
13021 ggtatttctg gaaaactgaa actgtttttc ctctactctg ataccacaag aatcatcagc
13081 acagaggaag acttctgtga tcaaatgtgg tgggagaggg aggttttcac cagcacatga
13141 gcagtcagtt ctgccgcaga ctcggcgggt gtccttcggt tcagttccaa caccgcctgc
13201 ctggagagag gtcagaccac agggtgaggg ctcagtcccc aagacataaa cacccaagac
13261 ataaacaccc aacaggtcca ccccgcctgc tgcccaggca gagccgattc accaagacgg
13321 gaattaggat agagaaagag taagtcacac agagccggct gtgcgggaga acggagttct
13381 attatgactc aaatcagtct ccccaagcat tcggggatca gagtttttaa ggataactta
13441 gtgtgtaggg ggccagtgag ttggagatga aagcgtaggg agtcgaaggt gtccttttgc
13501 gccgagtcag ttcctgggtg ggggccacaa gatcggatga gccagtttat caatccgggg
13561 gtgccagctg atccatggag tgcagggtct gcaaaatatc tcaagcactg attgatctta
13621 ggttttacaa tagtgatgtt accccaggaa caatttgggg aaggtcagaa tcttgtagcc
13681 tgtagctgca tgactcctaa accataattt cttttttgtt tttttttttt tatttttgag
13741 acagggtctc actctgtcac ctaggctgga gtgcagtggt gcaatcacag ctcactgcag
13801 cctcaacgtc gtaagctcaa gcgatcctcc cacctcagcc tgcctggtag ctgagactac
13861 aagcgacgcc ccagttaatt tttgtatttt tggtagaggc agcgttttgc cgtgtggccc
13921 tggctggtct cgaactcctg ggctcaagtg atccagcctc agcctcccaa agtgctggga
13981 caaccggggc cagtcactgc acctggccct aaaccataat ttctaatctt ttggctaatt
14041 tgttagtcct acaaaggcag tctagtcccc aggcaaaaag ggggtttgtt tcgggaaagg
14101 gctgttactg tctttgtttc aaactataaa ctaagttcct cctaaactta gttcggccta
14161 cacccaggaa tgaacaagga gagcttggag gttagaagca cgatggaatt ggttaggtca
14221 gatctctttc actgtctgag ttataatttt gcaatggtgg ttcaaagact gcccgcttct
14281 gacaccagtc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
14341 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
14401 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
14461 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
14521 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
14581 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
14641 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
14701 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
14761 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
14821 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
14881 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
14941 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
15001 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
15061 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
15121 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
15181 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
15241 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
15301 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
15361 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
15421 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
15481 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
15541 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
15601 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
15661 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
15721 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
15781 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
15841 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
15901 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
15961 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
16021 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
16081 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
16141 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
16201 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
16261 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
16321 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
16381 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
16441 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
16501 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
16561 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
16621 ggcgatcggt gcgggcctct tcgctattac gccaggggag gcagagattg cagtaagctg
16681 agatcgcagc actgcactcc agcctgggcg acagagtaag actctgtctc aaaaataaaa
16741 taaataaatc aatcagatat tccaatcttt tcctttattt atttatttat tttctatttt
16801 ggaaacacag tccttcctta ttccagaatt acacatatat tctatttttc tttatatgct
16861 ccagtttttt ttagaccttc acctgaaatg tgtgtataca aaatctaggc cagtccagca
16921 gagcctaaag gtaaaaaata aaataataaa aaataaataa aatctagctc actccttcac
16981 atcaaaatgg agatacagct gttagcatta aataccaaat aacccatctt gtcctcaata
17041 attttaagcg cctctctcca ccacatctaa ctcctgtcaa aggcatgtgc cccttccggg
17101 cgctctgctg tgctgccaac caactggcat gtggactctg cagggtccct aactgccaag
17161 ccccacagtg tgccctgagg ctgccccttc cttctagcgg ctgcccccac tcggctttgc
17221 tttccctagt ttcagttact tgcgttcagc caaggtctga aactaggtgc gcacagagcg
17281 gtaagactgc gagagaaaga gaccagcttt acagggggtt tatcacagtg caccctgaca
17341 gtcgtcagcc tcacaggggg tttatcacat tgcaccctga cagtcgtcag cctcacaggg
17401 ggtttatcac agtgcaccct tacaatcatt ccatttgatt cacaattttt ttagtctcta
17461 ctgtgcctaa cttgtaagtt aaatttgatc agaggtgtgt tcccagaggg gaaaacagta
17521 tatacagggt tcagtactat cgcatttcag gcctccacct gggtcttgga atgtgtcccc
17581 cgaggggtga tgactacctc agttggatct ccacaggtca cagtgacaca agataaccaa
17641 gacacctccc aaggctacca caatgggccg ccctccacgt gcacatggcc ggaggaactg
17701 ccatgtcgga ggtgcaagca cacctgcgca tcagagtcct tggtgtggag ggagggacca
17761 gcgcagcttc cagccatcca cctgatgaac agaacctagg gaaagcccca gttctactta
17821 caccaggaaa ggc
//
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NL4-3 mCherry Luciferase质粒载体应用举例
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