| 质粒名称: | pcDNA3.1 EYFP H148Q/I152L |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 25872 |
| 存储实验室: | Peter Haggie |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5400 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | EYFP H148Q/I152L |
| 物种来源: | A. victoria |
| 插入大小: | 700 bp |
| 基因突变: | EYFP from pEYFP (Clontech) with H148Q and I152L mutations |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HindIII (未破坏) |
| 3' 克隆位点: | EcoRI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | BGHrev |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3.1 EYFP H148Q/I152L was a gift from Peter Haggie (Addgene plasmid # 25872 ; http://n2t.net/addgene:25872 ; RRID:Addgene_25872) |
| 参考文献部分: | Green fluorescent protein-based halide indicators with improved chloride and iodide affinities. Galietta LJ, Haggie PM, Verkman AS. FEBS Lett. 2001 Jun 22. 499(3):220-4. 10.1016/S0014-5793(01)02561-3 PubMed 11423120 |
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 pcDNA3.1 EYFP H148Q/I152L 6181 bp ds-DNA circular SYN 29-JAN-2019
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1 EYFP H148Q/I152L
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6181)
AUTHORS Galietta LJ, Haggie PM, Verkman AS
TITLE Green fluorescent protein-based halide indicators with improved
chloride and iodide affinities.
JOURNAL FEBS Lett. 2001 Jun 22. 499(3):220-4.
PUBMED 11423120
REFERENCE 2 (bases 1 to 6181)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 29, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6181
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 726..735
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 732..1451
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSQNVYLMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(777..798)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1038..1057)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1385..1406
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1538..1555)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1544..1768
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1814..2242
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1901..1920)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2111..2132
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2251..2271)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2256..2585
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2436..2571
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2498..2517
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2652..3446
/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(2706..2725)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3316..3335
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3620..3741
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3657..3676)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3711..3730
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3790..3806)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3790..3806)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3803..3825)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3814..3830
/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(3838..3868)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3883..3904
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4021..4038)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4192..4780)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4272..4291)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4951..5811)
/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 5574..5593
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5812..5916)
/gene="bla"
/label=AmpR promoter
primer_bind complement(5991..6010)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
661 gctagcgttt aaacttaagc ttgcatgcct gcaggtcgac tctagaggat ccccgggtac
721 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
781 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
841 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
901 cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg
961 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
1021 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
1081 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
1141 tcctggggca caagctggag tacaactaca acagccagaa cgtctatctg atggccgaca
1201 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
1261 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
1321 ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg
1381 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
1441 tgtacaagta aagcggccgc gactctagaa ttctgcagat atccagcaca gtggcggccg
1501 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
1561 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
1621 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
1681 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
1741 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
1801 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
1861 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
1921 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
1981 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
2041 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
2101 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
2161 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
2221 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
2281 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
2341 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
2401 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
2461 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
2521 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
2581 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
2641 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
2701 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
2761 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
2821 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
2881 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
2941 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
3001 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
3061 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
3121 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
3181 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
3241 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
3301 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
3361 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
3421 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
3481 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
3541 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
3601 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
3661 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
3721 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
3781 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
3841 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
3901 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
3961 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
4021 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
4081 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
4141 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
4201 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
4261 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
4321 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
4381 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
4441 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
4501 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
4561 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
4621 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
4681 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
4741 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
4801 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
4861 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
4921 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
4981 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
5041 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
5101 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
5161 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
5221 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
5281 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
5341 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
5401 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
5461 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
5521 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
5581 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
5641 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
5701 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
5761 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
5821 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
5881 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
5941 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
6001 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
6061 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
6121 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
6181 t
//
