| 质粒名称: | pCX-ThPDE5VV |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 134778 |
| 存储实验室: | Xavier Nicol |
|
载体骨架基本信息 |
|
| 载体名称: | pCX |
| 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: | -- |
| 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: | ThPDE5VV |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | CAG |
| 标签/融合蛋白: |
MTURQOUISE2（插入时的N端子）
CPVenus-Venus（插入时的C端子）
|
| 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: | pCX-ThPDE5VV was a gift from Xavier Nicol (Addgene plasmid # 134778 ; http://n2t.net/addgene:134778 ; RRID:Addgene_134778) |
| 参考文献部分: | SponGee: A Genetic Tool for Subcellular and Cell-Specific cGMP Manipulation. Ros O, Zagar Y, Ribes S, Baudet S, Loulier K, Couvet S, Ladarre D, Aghaie A, Louail A, Petit C, Mechulam Y, Lenkei Z, Nicol X. Cell Rep. 2019 Jun 25;27(13):4003-4012.e6. doi: 10.1016/j.celrep.2019.05.102. 10.1016/j.celrep.2019.05.102 PubMed 31242429 |
买家导航
pCXThPDE5VV载体基本信息
pCXThPDE5VV载体图谱质粒图谱和多克隆位点信息
pCXThPDE5VV载体简介
pCXThPDE5VV载体序列
LOCUS pCX-ThPDE5VV 7598 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION cGMP FRET biosensor.
ACCESSION .
VERSION .
KEYWORDS pCX-ThPDE5VV
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7598)
AUTHORS Ros O, Zagar Y, Ribes S, Baudet S, Loulier K, Couvet S, Ladarre D,
Aghaie A, Louail A, Petit C, Mechulam Y, Lenkei Z, Nicol X
TITLE SponGee: A Genetic Tool for Subcellular and Cell-Specific cGMP
Manipulation.
JOURNAL Cell Rep. 2019 Jun 25;27(13):4003-4012.e6. doi:
10.1016/j.celrep.2019.05.102.
PUBMED 31242429
REFERENCE 2 (bases 1 to 7598)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7598
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 385..660
/label=chicken beta-actin promoter
intron 661..1677
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1601..1623
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1685..1704
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 2985..3719
/codon_start=1
/product="circularly permuted form of the Venus yellow
fluorescent protein (Nagai et al., 2004)"
/label=cp173Venus
/note="efficient FRET acceptor for CFP"
/translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
VYITADKQKNGIKANFKIRHNIE"
CDS 3777..4493
/codon_start=1
/product="yellow fluorescent protein (YFP) with fast and
efficient maturation (Nagai et al., 2002)"
/label=Venus
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4619..4638)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 4684..4739
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(4738..4757)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(5100..5116)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5100..5116)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5113..5135)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5124..5140
/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(5148..5178)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5193..5214
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5273..5468
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 5319..5454
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5381..5400
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 5474..5608
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5511..5530)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5565..5584
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5676..5693)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5847..6435)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5927..5946)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6606..7466)
/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 7229..7248
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7467..7571)
/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 tgggtggagt atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
481 cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg
541 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
1261 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt ctgataaact
1741 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca
1801 gcacagtggc ggccgcaatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
1861 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
1921 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
1981 ccgtgccctg gcccaccctc gtgaccaccc tgtcctgggg cgtgcagtgc ttcgcccgct
2041 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
2101 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
2161 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
2221 gcaacatcct ggggcacaag ctggagtaca actacatcag cgacaacgtc tatatcaccg
2281 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
2341 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
2401 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga
2461 agcgcgatca catggtcctg ctggagttcg tgaccgccgc ccgatgggaa ttagtgaagg
2521 atatttctag tcatttggat gtcacagcct tatgtcacaa aattttcttg catatccatg
2581 gactgatatc tgctgaccgc tattccctgt tccttgtctg tgaagacagc tccaatgaca
2641 agtttcttat cagccgcctc tttgatgttg ctgaaggttc aacactggaa gaagtttcaa
2701 ataactgtat ccgcttagaa tggaacaaag gcattgtggg acatgtggca gcgcttggtg
2761 agcccttgaa catcaaagat gcatatgagg atcctcggtt caatgcagaa gttgaccaaa
2821 ttacaggcta caagacacaa agcattcttt gtatgccaat taagaatcat agggaagagg
2881 ttgttggtgt agcccaggcc atcaacaaga aatcaggaaa cggtgggaca tttactgaaa
2941 aagatgaaaa ggactttgct gcttatttgg cagctagcga gctcatggac ggcggcgtgc
3001 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
3061 acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
3121 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
3181 acaagggtgg cagcggtggc atggtgagca agggcgagga gctgttcacc ggggtggtgc
3241 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
3301 gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc
3361 tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcctgcag tgcttcgccc
3421 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
3481 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
3541 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
3601 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
3661 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagc
3721 cgaattttgt cttcctgatc ggcgccgcag gaatactctt cgtatctagc gagctcatgg
3781 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
3841 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
3901 agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
3961 tgaccactct gggctacggc ctacagtgct tcgcccgcta ccccgaccac atgaagcagc
4021 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
4081 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
4141 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
4201 tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca
4261 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
4321 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
4381 tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
4441 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagccgaatt
4501 ccctcgaggt taacgcggcg cgcctctaat ctagcgaatt cactcctcag gtgcaggctg
4561 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
4621 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
4681 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
4741 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
4801 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
4861 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
4921 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
4981 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
5041 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
5101 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
5161 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
5221 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
5281 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
5341 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
5401 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
5461 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
5521 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
5581 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
5641 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
5701 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
5761 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
5821 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
5881 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
5941 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
6001 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
6061 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
6121 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
6181 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
6241 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
6301 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
6361 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
6421 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
6481 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
6541 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
6601 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
6661 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
6721 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
6781 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
6841 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
6901 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
6961 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
7021 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
7081 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
7141 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
7201 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
7261 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
7321 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
7381 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
7441 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
7501 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
7561 ggggttccgc gcacatttcc ccgaaaagtg ccacctgg
//
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pCXThPDE5VV载体质粒应用举例
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