| 质粒名称: | Double UP sfGFP to mScarlet |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 120261 |
| 存储实验室: | Erik Dent |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.4 |
| 完整质粒大小: | 6758 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | sfGFP/mScarletP2A-mcs |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 951 bp |
| 启动子: | CAG |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | CAGCTCCTGGGCAACGTGC |
| 3’测序引物 | m13R |
|
文献引用方法 |
|
| 材料和方法部分: |
Double UP sfGFP to mScarlet was a gift from Erik Dent (Addgene plasmid # 120261 ; http://n2t.net/addgene:120261 ; RRID:Addgene_120261)
|
| 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 sfGFP to mScarlet 6753 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Natively produces sfGFP, in the presence of Cre will be recombined
to produce mScarlet..
ACCESSION .
VERSION .
KEYWORDS Double UP sfGFP to mScarlet
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6753)
AUTHORS Taylor RJ, Carrington J, Gerlach LR, Taylor KL, Richters KE, Dent EW
TITLE Double UP: A Dual Color, Internally Controlled Platform for in utero
Knockdown or Overexpression.
JOURNAL Front Mol Neurosci. 2020 May 20;13:82. doi:
10.3389/fnmol.2020.00082. eCollection 2020.
PUBMED 32508591
REFERENCE 2 (bases 1 to 6753)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6753
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 5..384
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 663..1679
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1603..1625
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1687..1706
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
protein_bind complement(1740..1773)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS 1780..2499
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
primer_bind complement(1825..1846)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind 2433..2454
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(2503..2522)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 2568..2623
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
protein_bind complement(2939..2972)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS 2979..3674
/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(3112..3130)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3380..3399
/label=mCherry-F
/note="mCherry, forward primer"
CDS 3690..3746
/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_signal 3849..3904
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(4256..4272)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4256..4272)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4269..4291)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4280..4296
/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(4304..4334)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4349..4370
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4429..4624
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 4475..4610
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4537..4556
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 4630..4764
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4667..4686)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4721..4740
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4832..4849)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5003..5591)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5083..5102)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5762..6622)
/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 6385..6404
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6623..6727)
/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 gggggnsgcg cgccrggsgg ggsggggsgg ggsgrggggs
541 ggggsggggs grggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgctgcaga
1741 taacttcgta taatgtatgc tatacgaagt tatacgcgta tggtgagcaa gggcgaggag
1801 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
1861 ttcagcgtgc gcggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc
1921 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
1981 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc gccacgactt cttcaagtcc
2041 gccatgcccg aaggctacgt ccaggagcgc accatcagct tcaaggacga cggcacctac
2101 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
2161 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac
2221 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
2281 atccgccaca acgtggagga cggcagcgtg cagctcgccg accactacca gcagaacacc
2341 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgtg
2401 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
2461 gccgggatca ctcacggcat ggacgagctg tacaagtaaa gatctttttc cctctgccaa
2521 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
2581 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
2641 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
2701 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
2761 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
2821 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
2881 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctcat
2941 aacttcgtat aatgtatgct atacgaagtt atggtaccat ggtgagcaag ggcgaggcag
3001 tgatcaagga gttcatgcgg ttcaaggtgc acatggaggg ctccatgaac ggccacgagt
3061 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
3121 aggtgaccaa gggtggcccc ctgcccttct cctgggacat cctgtcccct cagttcatgt
3181 acggctccag ggccttcatc aagcaccccg ccgacatccc cgactactat aagcagtcct
3241 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gccgtgaccg
3301 tgacccagga cacctccctg gaggacggca ccctgatcta caaggtgaag ctccgcggca
3361 ccaacttccc tcctgacggc cccgtaatgc agaagaagac aatgggctgg gaagcgtcca
3421 ccgagcggtt gtaccccgag gacggcgtgc tgaagggcga cattaagatg gccctgcgcc
3481 tgaaggacgg cggacgctac ctggcggact tcaagaccac ctacaaggcc aagaagcccg
3541 tgcagatgcc cggcgcctac aacgtcgacc gcaagttgga catcacctcc cacaacgagg
3601 actacaccgt ggtggaacag tacgaacgct ccgagggccg ccactccacc ggcggcatgg
3661 acgagctgta caagttcgaa ggcagcggcg ctactaactt cagcctgctg aagcaggctg
3721 gagacgtgga agagaatcct ggacctatcg ataccggtca attggcggcc gcgcatgcgc
3781 tagctaattt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
3841 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
3901 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
3961 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
4021 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
4081 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
4141 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
4201 catagctgtc cctcttctct tatggagatc cctcgacaag cttggcgtaa tcatggtcat
4261 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
4321 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
4381 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta
4441 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
4501 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
4561 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
4621 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
4681 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
4741 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
4801 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
4861 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
4921 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
4981 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
5041 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
5101 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
5161 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
5221 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
5281 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
5341 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
5401 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
5461 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
5521 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
5581 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
5641 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
5701 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
5761 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
5821 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
5881 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
5941 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
6001 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
6061 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
6121 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
6181 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
6241 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
6301 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
6361 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
6421 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
6481 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
6541 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
6601 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
6661 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
6721 ttccgcgcac atttccccga aaagtgccac ctg
//
