| 质粒名称: | pCAGGS DRR mCherry Donor EF1a BFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 98896 |
| 存储实验室: | Jan Karlseder |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGGS |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3050 bp |
| 完整质粒大小: | 6435 bp |
| 载体修饰: | 删除 CBA 启动子 |
| 载体类型: | 哺乳动物表达，合成生物学 |
| 筛选标记: | BFP |
| 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: | Neomycin-T2A-mCherry-GFP |
| 物种来源: | Synthetic |
| 插入大小: | 2290 bp |
| 基因突变: | -- |
| 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: | pCAGGS DRR mCherry Donor EF1a BFP was a gift from Jan Karlseder (Addgene plasmid # 98896 ; http://n2t.net/addgene:98896 ; RRID:Addgene_98896) |
| 参考文献部分: | Regulation of DNA repair pathway choice in S and G2 phases by the NHEJ inhibitor CYREN. Arnoult N, Correia A, Ma J, Merlo A, Garcia-Gomez S, Maric M, Tognetti M, Benner CW, Boulton SJ, Saghatelian A, Karlseder J. Nature. 2017 Sep 20;549(7673):548-552. doi: 10.1038/nature24023. 10.1038/nature24023 PubMed 28959974 |
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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 pCAGGS DRR mCherry Donor EF1a BFP 6436 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Homologous recombination donor for pDRR. Expresses mCherry after
repait by HR..
ACCESSION .
VERSION .
KEYWORDS pCAGGS DRR mCherry Donor EF1a BFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6436)
AUTHORS Arnoult N, Correia A, Ma J, Merlo A, Garcia-Gomez S, Maric M,
Tognetti M, Benner CW, Boulton SJ, Saghatelian A, Karlseder J
TITLE Regulation of DNA repair pathway choice in S and G2 phases by the
NHEJ inhibitor CYREN.
JOURNAL Nature. 2017 Sep 20;549(7673):548-552. doi: 10.1038/nature24023.
PUBMED 28959974
REFERENCE 2 (bases 1 to 6436)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6436
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(87..103)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(87..103)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(100..122)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 111..127
/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(135..165)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 180..201
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 260..455
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 306..441
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 461..595
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(498..517)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 552..571
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(663..680)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(834..1422)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(914..933)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1593..2453)
/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 2216..2235
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2454..2558)
/gene="bla"
/label=AmpR promoter
CDS 2615..3412
/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="GSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2675..2694)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3285..3304
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
CDS 3413..3466
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 4183..4896
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
promoter 5024..5235
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 5298..5999
/codon_start=1
/product="monomeric blue fluorescent protein"
/label=TagBFP
/note="mammalian codon-optimized"
/translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
DLPSKLGHKLN"
primer_bind complement(6042..6061)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 6107..6162
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(6161..6180)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
ORIGIN
1 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
61 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
121 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
181 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
241 ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc
301 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
361 ttttttttat ttatacagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
421 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta
481 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
541 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg
601 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
661 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
721 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
781 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
841 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
901 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
961 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
1021 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
1081 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
1141 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
1201 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
1261 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
1321 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
1381 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
1441 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
1501 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
1561 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
1621 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
1681 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
1741 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
1801 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
1861 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
1921 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
1981 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
2041 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
2101 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
2161 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
2221 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
2281 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
2341 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
2401 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
2461 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
2521 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
2581 cctggtcgac attgattagt ccgactctca gtacggatcg gccattgaac aagatggatt
2641 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
2701 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
2761 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
2821 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
2881 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
2941 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
3001 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
3061 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
3121 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
3181 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
3241 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
3301 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
3361 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tcgagggcag
3421 aggaagtctt ctaacatgcg gtgacgtgga ggagaatccg ggccccatta gggtctctaa
3481 aggagaggag gataatatgg caattatcaa ggagtttatg agatttaagg tgcacatgga
3541 aggctccgtg aacggccacg agttcgagat tgagggagag ggcgagggac ggccctatga
3601 gggaacacag accgccaagc tgaaggtcac caagggagga cccctgcctt tcgcatggga
3661 catcctttca cctcagttca tgtacgggag taaagcgtat gtaaaacatc cagccgatat
3721 tcccgactat cttaaactgt cctttcccga aggttttaag tgggagagag tgatgaattt
3781 tgaagatggg ggcgttgtga cggttacaca ggattcttca ctccaagatg gcgagtttat
3841 atacaaggtt aagctgcgag gcacaaattt tccttccgat ggcccagtca tgcagaaaaa
3901 aactatgggg tgggaggcaa gctcagagcg aatgtacccc gaagatgggg ccctgaaagg
3961 agaaattaag caaagactga aactgaagga tggcggacac tacgacgccg aggtcaagac
4021 aacgtataag gccaaaaaac ctgtgcagct cccgggcgcc tataacgtga atatcaagct
4081 tgacataacc tctcacaacg aagactacac tatcgtggag cagtacgagc gcgctgaggg
4141 aaggcatagc accggaggga tggacgagtt gtataagtga gggtgagcaa gggcgaagaa
4201 ctctttacag gcgtcgttcc catcttggtt gagcttgacg gggacgtcaa cggccataaa
4261 ttcagcgttt ccggggaggg agagggcgat gcaacatacg ggaagctgac tttgaagttt
4321 atctgtacga ctggaaaact gccggtgcct tggccgacgc tggtgactac tctgacatat
4381 ggtgtgcagt gtttttccag atatcctgat catatgaagc agcatgattt cttcaagagc
4441 gcgatgccag agggctacgt gcaggaaaga acgatcttct tcaaagacga tggtaattac
4501 aaaactcggg cagaggtcaa attcgaggga gatacgctgg taaaccggat cgagctgaag
4561 ggcatcgatt ttaaggaaga tggtaacatc ctgggacaca aactggagta taattataac
4621 tcccataatg tctatattat ggctgacaag cagaagaacg ggatcaaggt gaactttaag
4681 atccggcaca atatcgaaga tggctccgtt cagctggccg atcattatca acaaaacaca
4741 cctattggag atgggccagt cctgttgcca gataatcact atctgtccac acagtccgcc
4801 ttgtccaaag acccgaacga gaagagagac cacatggtgc tgctggagtt tgttacggcg
4861 gccgggatca cgttgggcat ggatgagctt tacaagtgaa gcggccgcga ctcataatca
4921 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
4981 acctgaaaca taaaatgaat gcaattgaat caacctctgg attgggcaga gcgcacatcg
5041 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
5101 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
5161 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
5221 gccgccagaa cacagctctg gctaactaga gaacccactg tagtgaaccg tcagatcgcc
5281 tggagacgcc atccacgatg tccgagctga ttaaggagaa catgcacatg aagctgtaca
5341 tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct
5401 acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct
5461 tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg
5521 gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca
5581 catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc
5641 tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga
5701 agaaaacact cggctgggag gccttcaccg agacgctgta ccccgctgac ggcggcctgg
5761 aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacatca
5821 agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg
5881 tggactacag actggaaaga atcaaggagg ccaacaacga gacctacgtc gagcagcacg
5941 aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag cttaattgaa
6001 tctcatgctg gagttgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa
6061 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
6121 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
6181 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
6241 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
6301 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
6361 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
6421 ctagccagat ttttcc
//
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