质粒名称: | PB-CAG-BGHpA |
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出品公司: | Addgene |
目录编号: | 92161 |
存储实验室: | Xiaohua Shen |
载体骨架基本信息 |
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载体名称: | PiggyBac |
载体出品公司: | -- |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
筛选标记: | 潮霉素 |
载体抗性: | 氨苄霉素100 μg/mL |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | -- |
物种来源: | -- |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子: | -- |
标签/融合蛋白: | -- |
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 |
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材料和方法部分: | PB-CAG-BGHpA was a gift from Xiaohua Shen (Addgene plasmid # 92161 ; http://n2t.net/addgene:92161 ; RRID:Addgene_92161) |
参考文献部分: | Opposing Roles for the lncRNA Haunt and Its Genomic Locus in Regulating HOXA Gene Activation during Embryonic Stem Cell Differentiation. Yin Y, Yan P, Lu J, Song G, Zhu Y, Li Z, Zhao Y, Shen B, Huang X, Zhu H, Orkin SH, Shen X. Cell Stem Cell. 2015 May 7;16(5):504-16. doi: 10.1016/j.stem.2015.03.007. Epub 2015 Apr 16. 10.1016/j.stem.2015.03.007 PubMed 25891907 |
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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 PB-CAG-BGHpA 7531 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For lncRNA overexpression (in trans). ACCESSION . VERSION . KEYWORDS PB-CAG-BGHpA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7531) AUTHORS Yin Y, Yan P, Lu J, Song G, Zhu Y, Li Z, Zhao Y, Shen B, Huang X, Zhu H, Orkin SH, Shen X TITLE Opposing Roles for the lncRNA Haunt and Its Genomic Locus in Regulating HOXA Gene Activation during Embryonic Stem Cell Differentiation. JOURNAL Cell Stem Cell. 2015 May 7;16(5):504-16. doi: 10.1016/j.stem.2015.03.007. Epub 2015 Apr 16. PUBMED 25891907 REFERENCE 2 (bases 1 to 7531) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7531 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 383..658 /label=chicken beta-actin promoter intron 659..1667 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1591..1613 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind complement(1810..1827) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1816..2040 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" misc_feature 2125..2242 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind complement(2288..2321) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(2544..2563) /label=T7 /note="T7 promoter, forward primer" promoter complement(2545..2563) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2573..2590) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2573..2589) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2582..2604) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2731..3186 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2818..2837) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3028..3049 /label=F1ori-F /note="F1 origin, forward primer" promoter 3212..3316 /gene="bla" /label=AmpR promoter CDS 3317..4177 /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(3535..3554) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4348..4936 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4837..4856 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5090..5107 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5224..5245 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5260..5290 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5298..5314 /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 5303..5325 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5322..5338 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5322..5338 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 5357..5377 /label=T3 /note="T3 promoter, forward primer" promoter 5359..5377 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" repeat_region 5400..5434 /label=piggyBac left (5') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" CDS complement(5917..6942) /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" promoter complement(6961..7460) /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind 7402..7425 /label=MSCV-rev /note="Murine stem cell virus, reverse primer" protein_bind complement(7483..7516) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc 421 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg 481 atgggggcgg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg 541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc 601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg 661 agtcgctgcg ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg 721 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg 781 ctgtaattag cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct 841 taaagggctc cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt 901 gtgtgtgcgt ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg 961 gcgcggcgcg gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt 1021 gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg 1081 gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct 1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg 1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg 1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc 1321 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac 1381 ttcctttgtc ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag 1441 cgggcgcggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt 1501 gcgtcgccgc gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg 1561 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct 1621 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg 1681 tgctggttgt tgtgctgtct catcattttg gcaaagaatt cctcgacggt atcgataagc 1741 tatcacaagt ttgtacattc gaaggatcca cgcgtctcga gttaacggta cctctagaga 1801 attcgctagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg 1861 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa 1921 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca 1981 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg 2041 cttctgaggc ggttaattaa aataaaacta gagcgggccc ggtactaacc aaactggatc 2101 tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt tttgtaattc 2161 tttagtttgt atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc 2221 aatcccccct tttcttttaa aattgtggat gaatactgcc atttgtctgc agagcggccg 2281 cgtcgagata acttcgtata atgtatgcta tacgaagtta tgttttgtta ctttatagaa 2341 gaaattttga gtttttgttt ttttttaata aataaataaa cataaataaa ttgtttgttg 2401 aatttattat tagtatgtaa gtgtaaatat aataaaactt aatatctatt caaattaata 2461 aataaacgtc aattttacgc atgattatct ttaacgtacg tcacaatatg attatctttc 2521 tagggttaaa gagctccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg 2581 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag 2641 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc 2701 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg 2761 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 2821 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 2881 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 2941 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 3001 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 3061 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt 3121 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta 3181 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 3241 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 3301 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 3361 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 3421 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 3481 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 3541 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 3601 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 3661 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 3721 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 3781 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 3841 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga 3901 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 3961 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 4021 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 4081 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 4141 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 4201 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 4261 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 4321 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 4381 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 4441 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct 4501 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 4561 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 4621 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 4681 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 4741 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 4801 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 4861 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 4921 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 4981 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 5041 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 5101 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat 5161 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc 5221 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc 5281 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca 5341 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg gtaccttaac 5401 cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt 5461 ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc 5521 cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg 5581 agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat 5641 tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta 5701 tagatatcca actatgaaac attatcataa agcaatagta aagagccttc agtaaagagc 5761 aggcatttat ctaatcccac cccaccccca cccccgtagc tccaatcctt ccattcaaaa 5821 tgtaggtact ctgttctcac ccttcttaac aaagtatgac aggaaaaaac ttccatgtgg 5881 acatctttat tgtttaatag atcatcaatt tctcgactat tcctttgccc tcggacgagt 5941 gctggggcgt cggtttccac tatcggcgag tacttctaca cagccatcgg tccagacggc 6001 cgcgcttctg cgggcgattt gtgtacgccc gacagtcccg gctccggatc ggacgattgc 6061 gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca agctctgata 6121 gagttggtca agaccaatgc ggagcatata cgcccggagc cgcggcgatc ctgcaagctc 6181 cggatgcctc cgctcgaagt agcgcgtctg ctgctccata caagccaacc acggcctcca 6241 gaagaagatg ttggcgacct cgtattggga atccccgaac atcgcctcgc tccagtcaat 6301 gaccgctgtt atgcggccat tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac 6361 gaggtgccgg acttcggggc agtcctcggc ccaaagcatc agctcatcga gagcctgcgc 6421 gacggacgca ctgacggtgt cgtccatcac agtttgccag tgatacacat ggggatcagc 6481 aatcgcgcat atgaaatcac gccatgtagt gtattgaccg attccttgcg gtccgaatgg 6541 gccgaacccg ctcgtctggc taagatcggc cgcagcgatc gcatccatgg cctccgcgac 6601 cggctgcaga acagcgggca gttcggtttc aggcaggtct tgcaacgtga caccctgtgc 6661 acggcgggag atgcaatagg tcaggctctc gctgaattcc ccaatgtcaa gcacttccgg 6721 aatcgggagc gcggccgatg caaagtgccg ataaacataa cgatctttgt agaaaccatc 6781 ggcgcagcta tttacccgca ggacatatcc acgccctcct acatcgaagc tgaaagcacg 6841 agattcttcg ccctccgaga gctgcatcag gtcggagacg ctgtcgaact tttcgatcag 6901 aaacttctcg acagacgtcg cggtgagttc aggctttttc atggtggcgg ctggatcggt 6961 cgaaaggccc ggagatgagg aagaggagaa cagcgcggca gacgtgcgct tttgaagcgt 7021 gcgagaatgc cgggcctccg gaggaccttc gggcgcccgc cccgcccctg agcccgcccc 7081 tgagcccgcc cccggaccca ccccttccca gcctctgagc ccagaaagcg aaggagcaaa 7141 gctgctattg gccgctgccc caaaggccta cccgcttcca ttgctcagcg gtgctgtcca 7201 tctgcacgag actagtgaga cgtgctactt ccatttgtca cgtcctgcac gacgcgagct 7261 gcggggcggg ggggaacttc ctgactaggg gaggagtaga aggtggcgcg aaggggccac 7321 caaagaacgg agccggttgg cgcctaccgg tggatgtgga atgtgtgcga ggccagaggc 7381 cacttgtgta gcgccaagtg ccagcggggc tgctaaagcg catgctccag actgccttgg 7441 gaaaagcgcc tcccctaccc ggtagaattc atcgatgata tcataacttc gtataatgta 7501 tgctatacga agttatgcgg ccctcgaggt c //
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