| 质粒名称: | Double UP mNeon to mScarlet |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 125134 |
| 存储实验室: | Erik Dent |
|
载体骨架基本信息 |
|
| 载体名称: | pCAX |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6734 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: | -- |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | Removed Not1 site in mScarlet |
| 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: | Double UP mNeon to mScarlet was a gift from Erik Dent (Addgene plasmid # 125134 ; http://n2t.net/addgene:125134 ; RRID:Addgene_125134) |
| 参考文献部分: | Double UP: A Dual Color, Internally Controlled Platform for in utero Knockdown or Overexpression. Taylor RJ, Carrington J, Gerlach LR, Taylor KL, Richters KE, Dent EW. Front Mol Neurosci. 2020 May 20;13:82. doi: 10.3389/fnmol.2020.00082. eCollection 2020. 10.3389/fnmol.2020.00082 PubMed 32508591 |
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 Double UP mNeon to mScarlet 6737 bp ds-DNA circular SYN 17-MAY-2019
DEFINITION Natively produces sfGFP, in the presence of Cre will be recombined
to produce mScarlet..
ACCESSION .
VERSION .
KEYWORDS Double UP mNeon to mScarlet
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6737)
AUTHORS Taylor RJ, Carrington J, Taylor KL, Richters KE, Gerlach LR,, Dent
EW.
TITLE Double UP: efficient in utero electroporation via internal controls
JOURNAL BioRxiv 571323; doi: https://doi.org/10.1101/571323
REFERENCE 2 (bases 1 to 6737)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 17, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6737
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 5..384
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 662..1678
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1602..1624
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1686..1705
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
protein_bind complement(1739..1772)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS 1773..2483
/codon_start=1
/product="bright monomeric yellow-green fluorescent protein
derived from LanYFP (Shaner et al., 2013)"
/label=mNeonGreen
/translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
KAFTDVMGMDELYK"
regulatory 2028..2037
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind complement(2487..2506)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 2552..2607
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
protein_bind complement(2923..2956)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS 2963..3658
/codon_start=1
/product="bright monomeric red fluorescent protein evolved
from a synthetic template (Bindels et al., 2016)"
/label=mScarlet-I
/note="matures faster than mScarlet"
/translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
STGGMDELYK"
primer_bind complement(3096..3114)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3364..3383
/label=mCherry-F
/note="mCherry, forward primer"
CDS 3674..3730
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
polyA_site 3833..3888
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(4240..4256)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4240..4256)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4253..4275)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4264..4280
/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(4288..4318)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4333..4354
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4413..4608
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 4459..4594
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4521..4540
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 4614..4748
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4651..4670)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4705..4724
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4816..4833)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4987..5575)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5067..5086)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5746..6606)
/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 6369..6388
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6607..6711)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggsgcgc gccrggsggg gsggggsggg gsgrggggsg
541 gggsggggsg rggsggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
1261 ggggccgccy cgggccgggg rgggctcggg ggrggggcgc ggcggccccc ggagcgccgg
1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgctgcagat
1741 aacttcgtat aatgtatgct atacgaagtt atatggtgag caagggcgag gaggataaca
1801 tggcctctct cccagcgaca catgagttac acatctttgg ctccatcaac ggtgtggact
1861 ttgacatggt gggtcagggc accggcaatc caaatgatgg ttatgaggag ttaaacctga
1921 agtccaccaa gggtgacctc cagttctccc cctggattct ggtccctcat atcgggtatg
1981 gcttccatca gtacctgccc taccctgacg ggatgtcgcc tttccaggcc gccatggtag
2041 atggctccgg ctaccaagtc catcgcacaa tgcagtttga agatggtgcc tcccttactg
2101 ttaactaccg ctacacctac gagggaagcc acatcaaagg agaggcccag gtgaagggga
2161 ctggtttccc tgctgacggt cctgtgatga ccaactcgct gaccgctgcg gactggtgca
2221 ggtcgaagaa gacttacccc aacgacaaaa ccatcatcag tacctttaag tggagttaca
2281 ccactggaaa tggcaagcgc taccggagca ctgcgcggac cacctacacc tttgccaagc
2341 caatggcggc taactatctg aagaaccagc cgatgtacgt gttccgtaag acggagctca
2401 agcactccaa gaccgagctc aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg
2461 gcatggacga gctgtacaag taaagatctt tttccctctg ccaaaaatta tggggacatc
2521 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
2581 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
2641 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
2701 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
2761 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
2821 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
2881 ctcctctcct gactactccc agtcatagct gtccctcttc tcataacttc gtataatgta
2941 tgctatacga agttatggta ccatggtgag caagggcgag gcagtgatca aggagttcat
3001 gcggttcaag gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga
3061 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
3121 ccccctgccc ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt
3181 catcaagcac cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa
3241 gtgggagcgc gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc
3301 cctggaggac ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga
3361 cggccccgta atgcagaaga agacaatggg ctgggaagcg tccaccgagc ggttgtaccc
3421 cgaggacggc gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggacg
3481 ctacctggcg gacttcaaga ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc
3541 ctacaacgtc gaccgcaagt tggacatcac ctcccacaac gaggactaca ccgtggtgga
3601 acagtacgaa cgctccgagg gccgccactc caccggcggc atggacgagc tgtacaagtt
3661 cgaaggcagc ggcgctacta acttcagcct gctgaagcag gctggagacg tggaagagaa
3721 tcctggacct atcgataccg gtcaattggc ggccgcgcat gcgctagcta atttccctct
3781 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
3841 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
3901 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
3961 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
4021 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
4081 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
4141 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
4201 ctcttatgga gatccctcga caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
4261 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
4321 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
4381 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
4441 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
4501 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
4561 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
4621 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
4681 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
4741 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
4801 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
4861 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
4921 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
4981 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
5041 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
5101 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
5161 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
5221 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
5281 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
5341 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
5401 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
5461 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
5521 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
5581 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
5641 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
5701 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
5761 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
5821 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
5881 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
5941 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
6001 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
6061 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
6121 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
6181 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
6241 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
6301 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
6361 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
6421 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
6481 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
6541 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
6601 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
6661 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
6721 ccgaaaagtg ccacctg
//
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