| 质粒名称: | pCX-SponGee |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 134775 |
| 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: | SponGee,cGMP Sponge,SponGee-mRFP |
| 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: |
FLAG（插入件上的N端子）
mRFP（插件上的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-SponGee was a gift from Xavier Nicol (Addgene plasmid # 134775 ; http://n2t.net/addgene:134775 ; RRID:Addgene_134775) |
| 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
pCXSponGee载体基本信息
pCXSponGee载体图谱质粒图谱和多克隆位点信息
pCXSponGee载体简介
pCXSponGee载体序列
LOCUS pCX-SponGee 6633 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION mammalian expression of SponGee, a buffer of cGMP that prevents
downstream signaling.
ACCESSION .
VERSION .
KEYWORDS pCX-SponGee
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6633)
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 6633)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6633
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 385..659
/label=chicken beta-actin promoter
intron 660..1676
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1600..1622
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1684..1703
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1773..1782
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1782..1805
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
regulatory 2856..2865
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2862..3536
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/note="mammalian codon-optimized"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRPS
TGA"
primer_bind complement(2992..3010)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3260..3279
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 3454..3477
/label=DsRed1-C
/note="DsRed1, forward primer"
primer_bind complement(3654..3673)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 3719..3774
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(3773..3792)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(4135..4151)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4135..4151)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4148..4170)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4159..4175
/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(4183..4213)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4228..4249
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4308..4503
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 4354..4489
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4416..4435
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 4509..4643
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4546..4565)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4600..4619
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4711..4728)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4882..5470)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4962..4981)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5641..6501)
/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 6264..6283
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6502..6606)
/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 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
541 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
1261 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc tgatatcttt
1741 aattaaaccc cgggaccggt gctagctccg gagccaccat ggactacaaa gacgatgacg
1801 acaagtccgg aatcgatatg agcgagctgg aggaagactt tgccaagatt ctcatgctca
1861 aggaggagag gatcaaagag ctggagaagc ggctgtcaga gaaggaggaa gaaatccagg
1921 agctgaagag gaaactccat aaatgccagt cagtgctgcc cgtgccctcg acccacatcg
1981 gcccccggac cacccgggca cagggcatct cggccgagcc gcagacctac aggtccttcc
2041 acgacctccg agtgaccctg cccttctacc ccaagagtcc acagtccaag atcgatctca
2101 taaaggaggc catccttgac aatgacttta tgaagaactt ggagctgtca cagatccaag
2161 agattgtgga ttgtatgtac ccagtggagt acggcaaaga cagctgcatc atcaaagaag
2221 gagatgtggg gtcactggtg tatgtcatgg aagatggtaa ggttgaagtt acaaaagaag
2281 gcgtgaagct gtgcacaatg ggtcctggta aagtgtttgg agagttggct atcctttaca
2341 actgtacccg gacggcgacc gtcaaaactc ttgtaaatgt gaaactctgg gccattgatc
2401 gacaatgttt tcagacgata atgatgagga caggacttat caagcatacc gagtatatgg
2461 aatttttaaa aagcgttcca acattccaga gccttcctga agagatcctc agtaaacttg
2521 ctgacgtcct tgaagagacc cactatgaaa atggggaata tatcatcagg caaggtgcaa
2581 gaggggacac cttctttatc atcagtaaag gaaaggttaa tgtcactcgt gaagactcgc
2641 ccaatgaaga cccagtcttt cttagaacct taggaaaagg agattggttt ggagagaaag
2701 ccttgcaggg ggaagatgtg agaacagcga atgtaattgc ggcagaagct gtaacctgcc
2761 ttgtgatcga cagagactct ttcaaacatt tgattggagg attagatgat gtttctaata
2821 aagcatatga agatgcagaa gctaaggcgg atcccgccac catggcctcc tccgaggacg
2881 tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt
2941 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
3001 aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtcccct cagttccagt
3061 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
3121 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggatggcggc gtggtgaccg
3181 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
3241 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca
3301 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagatg aggctgaagc
3361 tgaaggacgg cggccactac gacgccgagg tcaagaccac ctacatggcc aagaagcccg
3421 tgcagctgcc cggcgcctac aagaccgaca tcaagctgga catcacctcc cacaacgagg
3481 actacaccat cgtggaacag tacgagcgcg ccgagggccg cccctccacc ggcgcctgta
3541 caagtaagaa ttctgcagat aaacgctagc gaattcactc ctcaggtgca ggctgcctat
3601 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
3661 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
3721 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
3781 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
3841 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
3901 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
3961 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
4021 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
4081 cctcttctct tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata
4141 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
4201 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
4261 ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag
4321 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
4381 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
4441 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
4501 aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
4561 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
4621 atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
4681 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
4741 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
4801 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
4861 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
4921 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
4981 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
5041 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
5101 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
5161 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
5221 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
5281 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
5341 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
5401 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
5461 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
5521 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
5581 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
5641 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
5701 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
5761 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
5821 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
5881 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
5941 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
6001 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
6061 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
6121 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
6181 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
6241 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
6301 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
6361 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
6421 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
6481 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
6541 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
6601 tccgcgcaca tttccccgaa aagtgccacc tgg
//
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