| 质粒名称: | CMV-ER-LAR-GECO1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 61244 |
| 存储实验室: | Robert Campbell |
|
载体骨架基本信息 |
|
| 载体名称: | Customized Vector |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3200 bp |
| 完整质粒大小: | 4514 bp |
| 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: | LAR-GECO1,low affinity red intensiometric genetically encoded Ca2+-indicators for optical |
| 物种来源: | Synthetic |
| 插入大小: | 1248 bp |
| 基因突变: | Substitutions relative to R-GECO1: V51W/I113V/N356S/D363Y/D381Y/F395A/V411A/L415I |
| GenBank编号 | KP091742 |
| 启动子 | CMV |
| 标签/融合蛋白: |
ER-targeting sequence: MLLPVPLLLGLLGAAAD (N端插入)
ER-retention sequence: KDEL (插入端C端)
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamH1(未破坏) |
| 3' 克隆位点: | EcoR1(未破坏) |
| 5' 测序引物 | TAATACGACTCACTATAGGG |
| 3' 测序引物 | TAGAAGGCACAGTCGAGG |
|
文献引用方法 |
|
| 材料和方法部分: | CMV-ER-LAR-GECO1 was a gift from Robert Campbell (Addgene plasmid # 61244 ; http://n2t.net/addgene:61244 ; RRID:Addgene_61244) |
| 参考文献部分: | Red fluorescent genetically encoded Ca2+ indicators for use in mitochondria and endoplasmic reticulum. Wu J, Prole DL, Shen Y, Lin Z, Gnanasekaran A, Liu Y, Chen L, Zhou H, Chen SR, Usachev YM, Taylor CW, Campbell RE. Biochem J. 2014 Nov 15;464(1):13-22. doi: 10.1042/BJ20140931. 10.1042/BJ20140931 PubMed 25164254 |
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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 CMV-ER-LAR-GECO1 4517 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses LAR-GECO1 in the endoplasmic reticulum in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS CMV-ER-LAR-GECO1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4517)
AUTHORS Wu J, Prole DL, Shen Y, Lin Z, Gnanasekaran A, Liu Y, Chen L, Zhou
H, Chen SR, Usachev YM, Taylor CW, Campbell RE
TITLE Red fluorescent genetically encoded Ca2+ indicators for use in
mitochondria and endoplasmic reticulum.
JOURNAL Biochem J. 2014 Nov 15;464(1):13-22. doi: 10.1042/BJ20140931.
PUBMED 25164254
REFERENCE 2 (bases 1 to 4517)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4517
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_signal 42..266
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 255..272
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
promoter 292..310
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 292..309
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(850..869)
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind complement(1721..1740)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1722..1740)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(1785..1988)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(1814..1834)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 1989..2368
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 2540..2559
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 2634..2738
/gene="bla"
/label=AmpR promoter
CDS 2739..3599
/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 complement(2957..2976)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3770..4358
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4259..4278
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ttccggacta gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc
61 cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
121 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
181 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
241 agatggctgg caactagaag gcacagtcga ggctgatcag cgagctctag catttaggtg
301 acactataga atagggccct ctagatgcat gctcgagcgg ccgccagtgt gatggatatc
361 tgcagaattc ttacagctcg tccttcttcg ctgtcatcat ttgtacaaac tcttcgtagt
421 ttacctgacc atccccatcg atgtctgcta ccctgatcat ttcatcaacc tcctcatctg
481 ttaacttctc tccaatgtct gtcatcgcgt ggcgaagctc tgctgcgccg atgtagccat
541 tgccgtcctt atccgccacg cggaacgctt ctctaatttc ctcttcactg tctgtgtaat
601 tcatttttct tgccatcatc gtcaggaact cagggaagtc gaatgtgccg tcaccgtcgg
661 catctacttc actgatcatg tcctgcagct ctgcttctgt ggggttctgc cccagagacc
721 gcatcaccgt ccccagctcc ttggttgtta tcgtcccatc cccgtccttg tcaaataggg
781 agaaagcctc tttaaattct gcgatctgct cttcagtcag ttggtcgcgc gtagcctccc
841 agcccatggt cttcttctgc attacggggc cgtcgggggg gaagttggtg ccgcgcagct
901 tcaccttgta gatgaatacg ccgtcctgca gggaggagtc ctggttaacg tgaataatgc
961 cgccgtcctc gaagttcatc acgcgctccc acctgaagcc ctcggggaag gacagcttga
1021 agtagtcggg gatgtcggct gggtgcttaa tgtaggcctt ggagccgtac atgaactgag
1081 gggacaggat gtcccaggcg aagggcaggg ggccaccctt ggtcaccttc agcttagcgg
1141 tctgaaaggc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt
1201 tcacggagcc ctccatgtgc accttgaagc gcatgaactc cttgatgatg gccatgttat
1261 cctcctcgcc cttgctcacc agactcccgc ctgtacctcc cttgtacagc tcgtccatgc
1321 cgccggtgga gtggcggccc tcggcgcgtt cgcactgttc cacgatggtg tagtcctcgt
1381 tgtgggacac gatgtccaac ttgatgtcga cgacgtaggc gccgggcagc tgcacgggct
1441 tcttggcctt gtaggtggtc ttgacctcgg cggcgtagtg gccgccgtcc ttcagcctca
1501 gccccttctt gatctcgctc ttcagggctc cgtcctcggg gtacatccgc tcggaaacca
1561 cgggtgagct cagccgacct atagctctcc atgcgtgacc tgccttattc cacttacgac
1621 gtgaagagtc gacgtcggcg gcggcgccca gcaggcccag cagcaggggg acgggcagca
1681 gcatggcggt ggatccgagc tcggtaccaa gcttgggtct ccctatagtg agtcgtatta
1741 atttcgataa gccagtaagc agtgggttct ctagttagcc agagagctct gcttatatag
1801 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
1861 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
1921 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
1981 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
2041 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
2101 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
2161 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
2221 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
2281 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
2341 tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac atcgcgaagc
2401 agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt
2461 gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca agcagggagc
2521 agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagggga
2581 tcgggagatc tcccgatccg tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa
2641 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
2701 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
2761 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
2821 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
2881 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
2941 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
3001 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
3061 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
3121 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
3181 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
3241 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
3301 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
3361 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
3421 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
3481 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
3541 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
3601 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
3661 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
3721 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
3781 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
3841 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
3901 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
3961 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
4021 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
4081 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
4141 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
4201 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
4261 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
4321 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
4381 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
4441 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
4501 cgaccgagcg cagcgag
//
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