| 质粒名称: | pCAG HRP-TM |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 44441 |
| 存储实验室: | Alice Ting |
|
载体骨架基本信息 |
|
| 载体名称: | pCAG |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5508 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Horseradish peroxidase fused to the transmembrane domain of PDGFR |
| 物种来源: | Synthetic |
| 插入大小: | 1224 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CAG |
| 标签/融合蛋白: |
HA tag（插入件上的N端子）
myc tag（插入件上的C端子）
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | HindIII (未破坏) |
| 5' 测序引物 | pCAG-F |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG HRP-TM was a gift from Alice Ting (Addgene plasmid # 44441 ; http://n2t.net/addgene:44441 ; RRID:Addgene_44441) |
| 参考文献部分: | Proteomic Mapping of Mitochondria in Living Cells via Spatially Restricted Enzymatic Tagging. Rhee HW, Zou P, Udeshi ND, Martell JD, Mootha VK, Carr SA, Ting AY. Science. 2013 Jan 31. 10.1126/science.1230593 PubMed 23371551 |
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 pCAG HRP-TM 5508 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCAG HRP-TM
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5508)
AUTHORS Rhee HW, Zou P, Udeshi ND, Martell JD, Mootha VK, Carr SA, Ting AY
TITLE Proteomic Mapping of Mitochondria in Living Cells via Spatially
Restricted Enzymatic Tagging.
JOURNAL Science. 2013 Jan 31.
PUBMED 23371551
REFERENCE 2 (bases 1 to 5508)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5508
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/label=chicken beta-actin promoter
intron 662..1670
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1594..1616
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1678..1697
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1763..1772
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1769..1831
/codon_start=1
/product="leader sequence from mouse immunoglobulin kappa
light chain"
/label=Ig-kappa leader
/translation="METDTLLLWVLLLWVPGSTGD"
CDS 1832..1858
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 2810..2839
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2843..2989
/codon_start=1
/product="transmembrane domain from platelet derived growth
factor receptor beta"
/label=PDGFR-beta TM domain
/translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
KKPR"
primer_bind complement(3011..3027)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3011..3027)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3024..3046)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3035..3051
/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(3059..3089)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3104..3125
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3184..3379
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 3230..3365
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3292..3311
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 3385..3519
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3422..3441)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3476..3495
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3587..3604)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3758..4346)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3838..3857)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4517..5377)
/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 5140..5159
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5378..5482)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attctgcagt cgacggtacc
1741 gcgggcccgg gatccaccgg tcgccaccat ggagacagac acactcctgc tatgggtact
1801 gctgctctgg gttccaggtt ccactggtga ctatccatat gatgttccag attatgctgg
1861 ggcccagcag ttaaccccta cattctacga caatagctgt cccaacgtgt ccaacatcgt
1921 tcgcgacaca atcgtcaacg agctcagatc cgatcccagg atcgctgctt caatattacg
1981 tctgcacttc catgactgct tcgtgaatgg ttgcgacgct agcatattac tggacaacac
2041 caccagtttc cgcactgaaa aggatgcatt cgggaacgct aacagcgcca ggggctttcc
2101 agtgatcgat cgcatgaagg ctgccgttga gtcagcatgc ccacgaacag tcagttgtgc
2161 agacctgctg actatagctg cgcaacagag cgtgactctt gcaggcggac cgtcctggag
2221 agtgccgctc ggtcgacgtg actccctaca ggcattccta gatctggcca acgccaactt
2281 gcctgctcca ttcttcaccc tgccccagct gaaggatagc tttagaaacg tgggtctgaa
2341 tcgctcgagt gaccttgtgg ctctgtccgg aggacacaca tttggaaaga accagtgtag
2401 gttcatcatg gataggctct acaatttcag caacactggg ttacctgacc ccacgctgaa
2461 cactacgtat ctccagacac tgagaggctt gtgcccactg aatggcaacc tcagtgcact
2521 agtggacttt gatctgcgga ccccaaccat cttcgataac aagtactatg tgaatctaga
2581 ggagcagaaa ggcctgatac agagtgatca agaactgttt agcagtccaa acgccactga
2641 caccatccca ctggtgagaa gttttgctaa ctctactcaa accttcttta acgccttcgt
2701 ggaagccatg gaccgtatgg gtaacattac ccctctgacg ggtacccaag gccagattcg
2761 tctgaactgc agagtggtca acagcaactc tccgcggctg caggtcgacg aacaaaaact
2821 catctcagaa gaggatctga atgctgtggg ccaggacacg caggaggtca tcgtggtgcc
2881 acactccttg ccctttaagg tggtggtgat ctcagccatc ctggccctgg tggtgctcac
2941 catcatctcc cttatcatcc tcatcatgct ttggcagaag aagccacgtt agaagcttgg
3001 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
3061 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
3121 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga
3181 tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
3241 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
3301 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
3361 aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa
3421 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
3481 ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc
3541 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
3601 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
3661 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
3721 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
3781 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
3841 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
3901 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
3961 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
4021 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
4081 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
4141 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
4201 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
4261 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
4321 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
4381 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
4441 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
4501 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
4561 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
4621 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
4681 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
4741 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
4801 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
4861 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
4921 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
4981 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
5041 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
5101 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
5161 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
5221 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
5281 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
5341 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
5401 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
5461 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
