| 质粒名称: | Double UP mClover3 to mScarlet |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 120262 |
| 存储实验室: | Erik Dent |
|
载体骨架基本信息 |
|
| 载体名称: | pCAX |
| 完整质粒大小: | 6752bp |
| 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: | Clover3/mScarletP2A-mcs |
| 基因突变: | Removed Not1 site in mScarlet |
| 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 mClover3 to mScarlet was a gift from Erik Dent (Addgene plasmid # 120262 ; http://n2t.net/addgene:120262 ; RRID:Addgene_120262)
|
| 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
<img .png="" alt="Double" mclover3="" data-cke-saved-src="/uploads/载体图谱/Double UP mClover3 to mScarlet质粒图谱.png" src="/uploads/载体图谱/Double UP mClover3 to mScarlet质粒图谱.png" "="">
载体简介
载体序列
LOCUS Double UP mClover3 to mScarlet 6752 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Natively produces mClover3, in the presence of Cre will be
recombined to produce mScarlet..
ACCESSION .
VERSION .
KEYWORDS Double UP mClover3 to mScarlet
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6752)
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 6752)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6752
/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 1779..2498
/codon_start=1
/product="bright green-yellow fluorescent protein derived
from mClover2, with improved photostability (Bajar et al.,
2003)"
/label=mClover3
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
primer_bind complement(1824..1845)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind 2432..2453
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(2502..2521)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 2567..2622
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
protein_bind complement(2938..2971)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS 2978..3673
/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(3111..3129)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3379..3398
/label=mCherry-F
/note="mCherry, forward primer"
CDS 3689..3745
/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 3848..3903
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(4255..4271)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4255..4271)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4268..4290)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4279..4295
/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(4303..4333)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4348..4369
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4428..4623
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 4474..4609
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4536..4555
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 4629..4763
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4666..4685)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4720..4739
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4831..4848)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5002..5590)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5082..5101)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5761..6621)
/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 6384..6403
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6622..6726)
/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 rggcggagag 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 ggggccgcct cgggccgggg agggctcggg ggaggggcgc 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 atacgcgtat ggtgagcaag ggcgaggagc
1801 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
1861 tcagcgtccg cggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca
1921 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg
1981 gcgtggcctg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
2041 ccatgcccga aggctacgtc caggagcgca ccatctcttt caaggacgac ggtacctaca
2101 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
2161 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca
2221 gccactacgt ctatatcacg gccgacaagc agaagaactg catcaaggct aacttcaaga
2281 tccgccacaa cgttgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
2341 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagccat cagtccaagc
2401 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
2461 ccgggattac acatggcatg gacgagctgt acaagtaaag atctttttcc ctctgccaaa
2521 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
2581 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
2641 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
2701 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
2761 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
2821 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
2881 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctcata
2941 acttcgtata atgtatgcta tacgaagtta tggtaccatg gtgagcaagg gcgaggcagt
3001 gatcaaggag ttcatgcggt tcaaggtgca catggagggc tccatgaacg gccacgagtt
3061 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
3121 ggtgaccaag ggtggccccc tgcccttctc ctgggacatc ctgtcccctc agttcatgta
3181 cggctccagg gccttcatca agcaccccgc cgacatcccc gactactata agcagtcctt
3241 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg ccgtgaccgt
3301 gacccaggac acctccctgg aggacggcac cctgatctac aaggtgaagc tccgcggcac
3361 caacttccct cctgacggcc ccgtaatgca gaagaagaca atgggctggg aagcgtccac
3421 cgagcggttg taccccgagg acggcgtgct gaagggcgac attaagatgg ccctgcgcct
3481 gaaggacggc ggacgctacc tggcggactt caagaccacc tacaaggcca agaagcccgt
3541 gcagatgccc ggcgcctaca acgtcgaccg caagttggac atcacctccc acaacgagga
3601 ctacaccgtg gtggaacagt acgaacgctc cgagggccgc cactccaccg gcggcatgga
3661 cgagctgtac aagttcgaag gcagcggcgc tactaacttc agcctgctga agcaggctgg
3721 agacgtggaa gagaatcctg gacctatcga taccggtcaa ttggcggccg cgcatgcgct
3781 agctaatttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact
3841 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
3901 tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt
3961 tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga
4021 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
4081 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
4141 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
4201 atagctgtcc ctcttctctt atggagatcc ctcgacaagc ttggcgtaat catggtcata
4261 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
4321 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
4381 ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag
4441 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
4501 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
4561 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
4621 aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
4681 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
4741 atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
4801 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
4861 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
4921 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
4981 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
5041 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
5101 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
5161 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
5221 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
5281 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
5341 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
5401 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
5461 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
5521 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
5581 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
5641 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
5701 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
5761 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
5821 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
5881 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
5941 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
6001 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
6061 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
6121 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
6181 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
6241 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
6301 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
6361 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
6421 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
6481 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
6541 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
6601 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
6661 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
6721 tccgcgcaca tttccccgaa aagtgccacc tg
//
