| 质粒名称: | pcDNA-K-GECO1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 105865 |
| 存储实验室: | Robert Campbell |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 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: | -- |
| 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: | K-GECO1 |
| 物种来源: | Synthetic |
| 插入大小: | 1239 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 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: | pcDNA-K-GECO1 was a gift from Robert Campbell (Addgene plasmid # 105865 ; http://n2t.net/addgene:105865 ; RRID:Addgene_105865) |
| 参考文献部分: | A genetically encoded Ca(2+) indicator based on circularly permutated sea anemone red fluorescent protein eqFP578. Shen Y, Dana H, Abdelfattah AS, Patel R, Shea J, Molina RS, Rawal B, Rancic V, Chang YF, Wu L, Chen Y, Qian Y, Wiens MD, Hambleton N, Ballanyi K, Hughes TE, Drobizhev M, Kim DS, Koyama M, Schreiter ER, Campbell RE. BMC Biol. 2018 Jan 16;16(1):9. doi: 10.1186/s12915-018-0480-0. 10.1186/s12915-018-0480-0 PubMed 29338710 |
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 pcDNA-K-GECO1 6598 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Mammalian cell expression plasmid for red fluorescent Ca2+ indicator
K-GECO1.
ACCESSION .
VERSION .
KEYWORDS pcDNA-K-GECO1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6598)
AUTHORS Shen Y, Dana H, Abdelfattah AS, Patel R, Shea J, Molina RS, Rawal B,
Rancic V, Chang YF, Wu L, Chen Y, Qian Y, Wiens MD, Hambleton N,
Ballanyi K, Hughes TE, Drobizhev M, Kim DS, Koyama M, Schreiter ER,
Campbell RE
TITLE A genetically encoded Ca(2+) indicator based on circularly
permutated sea anemone red fluorescent protein eqFP578.
JOURNAL BMC Biol. 2018 Jan 16;16(1):9. doi: 10.1186/s12915-018-0480-0.
PUBMED 29338710
REFERENCE 2 (bases 1 to 6598)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6598
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 110..489
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 490..693
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 644..664
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 738..757
/label=T7
/note="T7 promoter, forward primer"
promoter 738..756
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 781..790
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind complement(2064..2081)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2070..2294
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2340..2768
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2427..2446)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2637..2658
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2777..2797)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2782..3111
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2962..3097
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3024..3043
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3178..3972
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3232..3251)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3842..3861
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4146..4267
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4183..4202)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4237..4256
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4316..4332)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4316..4332)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4329..4351)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4340..4356
/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(4364..4394)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4409..4430
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4547..4564)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4718..5306)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4798..4817)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5477..6337)
/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 6100..6119
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6338..6442)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6517..6536)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
61 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
361 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
421 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
481 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
541 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
601 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
661 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
721 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgtgcc
781 gccaccatgg cctcgagaat gggcagcgtg aagctgatcc ccagcctgac caccgtgatc
841 ctcgtgaagt ccatgctgcg gaagcggagc ttcggcaacc ccttcaagta taatacggag
901 accctgtacc ccgctgacgg cggcctggaa ggcgcatgtg acatggccct gaagctcgtg
961 ggcgggggcc acctgaactg cagccttgag accacataca gatccaagaa acccgctacg
1021 aacctcaaga tgcccggtgt ctacaacgtg gaccacagac tggaacgaat caaagaggcc
1081 gacgatgaga cctacgtcga gctgcacgag gtggctgtgg ccagatacgt gggcctgggt
1141 ggtggcggag gtacaggcgg gagtgtgagc gagctgatta aggagaacat gccaatgaag
1201 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc
1261 aagccctacg agggcaccca gaccatgaga atcaaggtcg tcgagggcgg ccctctcccc
1321 ttcgccttcg acatcctggc taccagcttc atgtacggca gcagaacctt catcaagcac
1381 cctcctggca tccccgactt ctttaagcag tccttccctg agggcttcac atgggagaga
1441 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
1501 ggctgcctca tctacaacgt caaggttaga gggatgaact tcccagccaa cggccctgtg
1561 atgcagaaga aaacactcgg ctgggaggcc agtaatggcc aactgactga agagcagatc
1621 gcagaattta aagaggcttt ctccctattt gacaaggacg gggatgggac gataacaacc
1681 aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc ggagctgcag
1741 gacatgatca atgaagtaga tgccgacggt gacggcacat tcgacttccc tgagttcctg
1801 acgatgatgg caagaaaaat gagttataga gtcactgaag aggaaattag agaagcgttc
1861 cgcgtgtttg ataaggacgg caatggctac atcggcgcag cagagcttcg ccacgtgatg
1921 acagaccttg gagagaagtt aacagatgag gaggttgatg aaatgatcag ggtagcagac
1981 atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac agcgaagtag
2041 aagcttggta aaccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
2101 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
2161 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
2221 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
2281 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
2341 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
2401 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
2461 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
2521 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
2581 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
2641 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
2701 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
2761 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
2821 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
2881 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
2941 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
3001 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
3061 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
3121 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
3181 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
3241 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
3301 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
3361 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
3421 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
3481 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
3541 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
3601 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
3661 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
3721 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
3781 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
3841 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
3901 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
3961 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
4021 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
4081 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
4141 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
4201 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
4261 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
4321 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
4381 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
4441 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
4501 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
4561 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
4621 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
4681 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
4741 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
4801 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
4861 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
4921 gctataggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
4981 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
5041 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
5101 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
5161 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
5221 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
5281 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
5341 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
5401 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
5461 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
5521 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
5581 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
5641 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
5701 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
5761 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
5821 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
5881 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
5941 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
6001 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
6061 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
6121 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
6181 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
6241 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
6301 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
6361 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
6421 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
6481 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
6541 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagc
//
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