| 质粒名称: | pCAG mito-GCaMP5G |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 105009 |
| 存储实验室: | Franck Polleux |
|
载体骨架基本信息 |
|
| 载体名称: | pCIG2 |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | GCaMP5G |
| 启动子: | CAG |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | 未知(未破坏) |
| 3’克隆位点: | 未知(未破坏) |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG mito-GCaMP5G was a gift from Franck Polleux (Addgene plasmid # 105009 ; http://n2t.net/addgene:105009 ; RRID:Addgene_105009) |
| 参考文献部分: | LKB1 Regulates Mitochondria-Dependent Presynaptic Calcium Clearance and Neurotransmitter Release Properties at Excitatory Synapses along Cortical Axons. Kwon SK, Sando R 3rd, Lewis TL, Hirabayashi Y, Maximov A, Polleux F. PLoS Biol. 2016 Jul 18;14(7):e1002516. doi: 10.1371/journal.pbio.1002516. eCollection 2016 Jul. PBIOLOGY-D-16-00343 [pii] PubMed 27429220 |
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 mito-GCaMP5G 6289 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION mitochondrial calcium sensor.
ACCESSION .
VERSION .
KEYWORDS pCAG mito-GCaMP5G
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6289)
AUTHORS Kwon SK, Sando R 3rd, Lewis TL, Hirabayashi Y, Maximov A, Polleux F
TITLE LKB1 Regulates Mitochondria-Dependent Presynaptic Calcium Clearance
and Neurotransmitter Release Properties at Excitatory Synapses along
Cortical Axons.
JOURNAL PLoS Biol. 2016 Jul 18;14(7):e1002516. doi:
10.1371/journal.pbio.1002516. eCollection 2016 Jul.
PUBMED 27429220
REFERENCE 2 (bases 1 to 6289)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6289
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(42..629)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(122..141)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(800..1660)
/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 1423..1442
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1661..1765)
/gene="bla"
/label=AmpR promoter
enhancer 1796..2175
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2177..2454
/label=chicken beta-actin promoter
intron 2455..3467
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 3391..3413
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 3475..3494
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 3528..3602
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label=COX8 presequence
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 3639..3656
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 3660..3692
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/label=T7 tag (gene 10 leader)
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS 3696..3719
/codon_start=1
/product="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/label=Xpress(TM) tag
/translation="DLYDDDDK"
regulatory 3726..3735
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3747..3803
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKTGHAVRAIGRLSS"
primer_bind 4016..4037
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(4143..4164)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4404..4423)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
CDS 4539..4982
/codon_start=1
/product="calcium-binding protein"
/label=calmodulin
/translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
ELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGNGYISAAEL
RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
primer_bind complement(5073..5092)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 5138..5193
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(5192..5211)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(5584..5600)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5584..5600)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5597..5619)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5608..5624
/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(5632..5662)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5677..5698
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5757..5952
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 5803..5938
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5865..5884
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 5958..6092
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5995..6014)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6049..6068
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6160..6177)
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
61 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
481 agacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
1021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
1081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
1141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
1201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
1261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
1321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
1381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
1441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
1501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
1561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
1621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
1681 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
1741 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcgacat
1801 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
1861 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
1921 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
1981 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
2041 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
2101 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
2161 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
2221 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
2281 gcgggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg
2341 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
2401 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
2461 gctgcgttgc cttcgccccg tgccccgctc cgcgccgcct cgcgccgccc gccccggctc
2521 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
2581 aattagcgct tggtttaatg acggctcgtt tcttttctgt ggctgcgtga aagccttaaa
2641 gggctccggg agggcccttt gtgcgggggg gagcggctcg gggggtgcgt gcgtgtgtgt
2701 gtgcgtgggg agcgccgcgt gcggcccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
2761 ggcgcggggc tttgtgcgct ccgcgtgtgc gcgaggggag cgcggccggg ggcggtgccc
2821 cgcggtgcgg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg
2881 gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac ccccctcccc
2941 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
3001 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
3061 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
3121 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
3181 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
3241 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
3301 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
3361 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
3421 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
3481 tgctggttat tgtgctgtct catcattttg gcaaagaatt gctcgagatg tccgtcctga
3541 cgccgctgct gctgcggggc ttgacaggct cggcccggcg gctcccagtg ccgcgcgcca
3601 agatccattc gttgggggat ccaccggtca tgggttctca tcatcatcat catcatggta
3661 tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg
3721 atctcgccac catggtcgac tcatcacgtc gtaagtggaa taagacaggt cacgcagtca
3781 gagctatagg tcggctgagc tcactcgaga acgtctatat caaggccgac aagcagaaga
3841 acggcatcaa ggcgaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
3901 cctaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
3961 actacctgag cgtgcagtcc aaactttcga aagaccccaa cgagaagcgc gatcacatgg
4021 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
4081 gcggtaccgg agggagcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
4141 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg
4201 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
4261 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
4321 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacatcc
4381 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
4441 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
4501 gcaacatcct ggggcacaag ctggagtaca acctgccgga ccaactgact gaagagcaga
4561 tcgcagaatt taaagaggct ttctccctat ttgacaagga cggggatggg acaataacaa
4621 ccaaggagct ggggacggtg atgcggtctc tggggcagaa ccccacagaa gcagagctgc
4681 aggacatgat caatgaagta gatgccgacg gtgacggcac aatcgacttc cctgagttcc
4741 tgacaatgat ggcaagaaaa atgaaataca cagacagtga agaagaaatt agagaagcgt
4801 tccgtgtgtt tgataaggat ggcaatggct acatcagtgc agcagagctt cgccacgtga
4861 tgacaaacct tggagagaag ttaacagatg aagaggttga tgaaatgatc agggaagcag
4921 acatcgatgg ggatggtcag gtaaactacg aagagtttgt acaaatgatg acagcgaagt
4981 gagcggccgc aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
5041 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
5101 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
5161 gcaatagtgt gttggaattt tttgtgtcta tcactcggaa ggacatatgg gagggcaaat
5221 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
5281 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
5341 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
5401 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
5461 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
5521 ctcgacctgc accgtcgacc agctggtcga cggtgcaccg tcgaccagct tggcgtaatc
5581 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
5641 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
5701 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat
5761 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
5821 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
5881 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
5941 ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata
6001 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
6061 aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc
6121 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
6181 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
6241 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccag
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：LentiCRISPRv2-mCherry
- 下一篇：pSLIK CA Rac1
