| 质粒名称: | ArcLight-A242 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 36857 |
| 存储实验室: | Vincent Pieribone |
|
载体骨架基本信息 |
|
| 载体名称: | pCS2+ |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5694 bp |
| 完整质粒大小: | 7146 bp |
| 载体修饰: | 插入一个抗潮溶素基因，以便于选择稳定转染的细胞。 |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 潮霉素 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | fluorescent protein voltage sensor,ArcLight,Ci-VSP,super ecliptic pHluorin |
| 物种来源: | Ciona intestinalis |
| 插入大小: | 1452 bp |
| 基因突变: | Ci-VSP contains R217Q mutation; super ecliptic pHluorin contains A227D mutation |
| GenBank编号 | AB183035 AY533296 |
| 启动子 | semian CMV |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HindIII(未破坏) |
| 3' 克隆位点: | XbaI (未破坏) |
| 5' 测序引物 | GACGTAAATGGGCGGTAGGCGTG |
| 3' 测序引物 | TTAAAAAACCTCCCACACCTC |
|
文献引用方法 |
|
| 材料和方法部分: | ArcLight-A242 was a gift from Vincent Pieribone (Addgene plasmid # 36857 ; http://n2t.net/addgene:36857 ; RRID:Addgene_36857) |
| 参考文献部分: | Single action potentials and subthreshold electrical events imaged in neurons with a fluorescent protein voltage probe. Jin L, Han Z, Platisa J, Wooltorton JR, Cohen LB, Pieribone VA. Neuron. 2012 Sep 6;75(5):779-85. 10.1016/j.neuron.2012.06.040 PubMed 22958819 |
买家导航
ArcLightA242载体基本信息
ArcLightA242载体图谱质粒图谱和多克隆位点信息
ArcLightA242载体简介
ArcLightA242载体序列
LOCUS ArcLight-A242 7143 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS ArcLight-A242
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7143)
AUTHORS Jin L, Han Z, Platisa J, Wooltorton JR, Cohen LB, Pieribone VA
TITLE Single action potentials and subthreshold electrical events imaged
in neurons with a fluorescent protein voltage probe.
JOURNAL Neuron. 2012 Sep 6;75(5):779-85.
PUBMED 22958819
REFERENCE 2 (bases 1 to 7143)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7143
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 35..53
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 35..52
/label=SP6
/note="SP6 promoter, forward primer"
CDS 799..1515
/codon_start=1
/product="pH-sensitive mutant of green fluorescent protein
(Ng et al., 2002)"
/label=superecliptic pHluorin
/note="excited strongly by blue light at neutral pH but
minimally fluorescent at acidic pH"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKA
NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLE
FVTADGITHGMDELYK"
primer_bind complement(829..857)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 1452..1472
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
promoter complement(1523..1540)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 1545..1679
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1784..1804)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 1789..2118
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 1969..2104
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2031..2050
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2167..3192
/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"
polyA_signal 3322..3443
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3481..3497)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3481..3497)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3494..3516)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3505..3521
/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(3529..3559)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3574..3595
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3712..3729)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3883..4471)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3963..3982)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4642..5502)
/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 5265..5284
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5503..5607)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5633..6088)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5770..5791)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5982..6001
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 6166..7143
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
ORIGIN
1 cgccattctg cctggggacg tcggagctag cttgatttag gtgacactat agaatacaag
61 cttatggagg gattcgacgg ttcagatttt agtcctccag ctgatttagt tggcgttgac
121 ggtgcagtca tgcggaacgt cgttgacgtc acgataaatg gtgacgtcac tgctccgccg
181 aaagctgcgc caagaaaaag tgaatcggta aagaaagttc attggaatga tgtagaccaa
241 ggaccgagtg aaaaaccaga gacaagacag gaggaacgaa tagatatacc cgagatttca
301 ggtctatggt ggggcgagaa tgaacatgga gtggacgatg ggagaatgga gatacctact
361 actggtgtag gtcgcgtcca gtttcgtgtc cgagcagtga ttgatcatct agggatgcga
421 gtctttggag tcttcctaat tttcttggac atcatcctca tgatcattga tctcagtctt
481 ccaggaaaaa gtgaatcttc acaatccttt tatgacggga tggctttggc tctttcttgt
541 tatttcatgc tggatttagg attaaggata tttgcctacg ggcccaagaa tttcttcacc
601 aacccctggg aggttgctga tggtttgatt atcgttgtca cattcgtcgt cacgatattt
661 tacactgtgt tagatgaata cgtgcaagaa acaggagccg atggtttggg gcaattggtt
721 gtgttggccc gtttgctgcg tgtggttcga ttagcaagaa tattttattc ccaccaacaa
781 atgaaggctg gggatcccat gagtaaagga gaagaacttt tcactggagt tgtcccaatt
841 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa
901 ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct
961 gttccatggc caacacttgt cactacttta acttatggtg ttcaatgctt ttcaagatac
1021 ccagatcata tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
1081 gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt
1141 gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga
1201 aacattcttg gacacaaatt ggaatacaac tataacgatc accaggtgta catcatggca
1261 gacaaacaaa agaatggaat caaagctaac ttcaaaatta gacacaacat tgaagatgga
1321 ggcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg gcccgtcctt
1381 ttaccagaca accattacct gtttacaact tctactcttt cgaaagatcc caacgaaaag
1441 agagaccaca tggtccttct tgagtttgta acagctgatg ggattacaca tggcatggat
1501 gaactataca aatgatctag aactatagtg agtcgtatta cgtagatcca gacatgataa
1561 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
1621 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
1681 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
1741 aattgatatc cgcggcgcca atgcattggg cccggtaccc tgtggaatgt gtgtcagtta
1801 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
1861 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
1921 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
1981 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
2041 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
2101 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
2161 cacgtgatga aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
2221 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
2281 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
2341 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
2401 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
2461 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
2521 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
2581 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
2641 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
2701 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
2761 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
2821 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
2881 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
2941 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
3001 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
3061 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
3121 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
3181 gcaaaggaat agcacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
3241 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
3301 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
3361 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
3421 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt acgcgcttgg cgtaatcatg
3481 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
3541 cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
3601 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
3661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
3721 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
3781 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
3841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
3901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
3961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
4021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
4081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
4141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
4201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
4261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
4321 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
4381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
4441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
4501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
4561 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
4621 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
4681 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
4741 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
4801 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
4861 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
4921 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
4981 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
5041 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
5101 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
5161 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
5221 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
5281 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
5341 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
5401 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
5461 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
5521 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
5581 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
5641 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
5701 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
5761 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
5821 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
5881 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
5941 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
6001 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
6061 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
6121 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg
6181 cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg
6241 cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg
6301 cggacttggc accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg
6361 gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat
6421 attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata
6481 taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt
6541 attggcccta tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc
6601 tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc atagccactc
6661 ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc
6721 ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc
6781 tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg
6841 cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg
6901 gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc
6961 agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc
7021 caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg
7081 acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg
7141 aac
//
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