质粒名称: | pLenti.PGK.chFP.W |
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出品公司: | 爱迪基因 |
目录编号: | 51008 |
存储实验室: | Rusty Lansford |
载体骨架基本信息 |
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载体名称: | pRRLsin.cPPT.PGKp.eGFP.W |
载体出品公司: | L Naldini |
载体抗性: | 氨苄青霉素 |
空载体大小: | -- |
完整质粒大小: | 7375 bp |
载体修饰: | pLenti.pgk:chFP慢病毒载体衍生自pRRLsin.cPPT.PGKp.eGFP.W慢病毒载体,其包含组成型表达的人磷酸甘油酸激酶启动子(De Palma等,2003)。 |
载体类型: | 哺乳细胞表达,慢病毒 |
筛选标记: | -- |
高拷贝/低拷贝: | 低拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | mChFP,mCherry FP |
物种来源: | -- |
插入大小: | 711 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | human PGK |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | BamH1(未破坏) |
3' 克隆位点: | Sal11(未破坏) |
5' 测序引物 | T3 |
3' 测序引物 | SV40pA-R |
文献引用方法 |
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材料和方法部分: | pLenti.PGK.chFP.W was a gift from Rusty Lansford (Addgene plasmid # 51008 ; http://n2t.net/addgene:51008 ; RRID:Addgene_51008) |
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 pLenti.PGK.chFP.W 7363 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Lentivector that expresses chFP from internal human PGK promoter. ACCESSION . VERSION . KEYWORDS pLenti.PGK.chFP.W SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7363) TITLE Lentivirus JOURNAL Unpublished REFERENCE 2 (bases 1 to 7363) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7363 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 236..416 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 460..585 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1078..1311 /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 1807..1924 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 1971..2481 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 2400..2418 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 2491..3201 /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(2636..2654) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2904..2923 /label=mCherry-F /note="mCherry, forward primer" misc_feature 3217..3805 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3270..3290) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3688..3699) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3892..4125 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4197..4318 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4234..4253) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4288..4307 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4337..4472 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4399..4418 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4504..4523) /label=T7 /note="T7 promoter, forward primer" promoter complement(4505..4523) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4533..4550) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4533..4549) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4542..4564) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4694..5149 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4781..4800) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4991..5012 /label=F1ori-F /note="F1 origin, forward primer" promoter 5178..5282 /gene="bla" /label=AmpR promoter CDS 5283..6143 /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(5501..5520) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6314..6902 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6803..6822 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7056..7073 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7190..7211 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7226..7256 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7264..7280 /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 7269..7291 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7288..7304 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7288..7304 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 7323..7343 /label=T3 /note="T3 promoter, forward primer" promoter 7325..7343 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag 61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg 121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact 181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacaa taaacgggtc 241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg 421 cgcccgaaca gggacctgaa agcgaaaggg aaaccagagc tctctcgacg caggactcgg 481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt 541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg 601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa 661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg 721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca 781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa 841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa 901 agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag 961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt 1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg 1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cctcaatgac 1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct 1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct 1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg 1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa 1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa 1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa 1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac 1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag 1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc 1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga 1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg 1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga 1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa 1921 ttttatcgat cacgagacta gcctcgagaa gcttgatatc gaattcccac ggggttgggg 1981 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt 2041 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc 2101 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc 2161 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca 2221 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg 2281 cgatgggctg tggccaatag cggctgctca gcggggcgcg ccgagagcag cggccgggaa 2341 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg 2401 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga 2461 atcaccgacc tctctcccca gggggatccg atggtgagca agggcgagga ggataacatg 2521 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac 2581 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag 2641 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc 2701 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg 2761 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg 2821 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc 2881 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc 2941 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg 3001 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag 3061 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac 3121 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc 3181 atggacgagc tgtacaagta aagcggccgc gtcgacaatc aacctctgga ttacaaaatt 3241 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 3301 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 3361 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 3421 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 3481 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 3541 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 3601 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg 3661 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 3721 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 3781 atctcccttt gggccgcctc cccgcctgga attcgagctc ggtaccttta agaccaatga 3841 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc 3901 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag 3961 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat 4021 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact 4081 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc 4141 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact 4201 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 4261 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 4321 atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 4381 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt 4441 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcgtcgaga cgtacccaat 4501 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac 4561 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc 4621 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat 4681 ggcgaatggc gcgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 4741 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 4801 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 4861 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 4921 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 4981 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 5041 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 5101 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc ccaggtggca 5161 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 5221 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 5281 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 5341 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 5401 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 5461 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 5521 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 5581 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 5641 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 5701 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 5761 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 5821 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 5881 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 5941 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 6001 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 6061 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 6121 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 6181 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 6241 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 6301 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 6361 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 6421 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt 6481 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 6541 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 6601 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 6661 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 6721 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 6781 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 6841 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 6901 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 6961 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag 7021 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg 7081 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct 7141 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt 7201 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg 7261 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc 7321 gcgcaattaa ccctcactaa agggaacaaa agctggagct gca //