| 质粒名称: | PB-CAG-BGHpA |
|---|---|
| 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 |
|
| 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 |
|
| 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 |
|
| 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 |
买家导航
PB-CAG-BGHpA载体质粒基本信息
PB-CAG-BGHpA质粒图谱载体图谱和PB-CAG-BGHpA载体序列质粒序列多克隆位点信息
PB-CAG-BGHpA质粒载体简介
PB-CAG-BGHpA质粒序列载体序列
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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