| 质粒名称: | pcDNA3.1+(Zeo)-FLAG-muGFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 166726 |
| 存储实验室: | Daniel Scott |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1+(Zeo) |
| 载体出品公司: | Clontech |
| 空载体大小: | 4985 bp |
| 完整质粒大小: | 5702 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | 博莱霉素 |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 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: | muGFP |
| 物种来源: | synthetic/engineered |
| 插入大小: | 711 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: | FLAG（插入件上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI (未破坏) |
| 3’克隆位点: | SpeI (未破坏) |
| 5’测序引物 | TGGCTAACTAGAGAACCCAC |
| 3’测序引物 | CCTACTCAGACAATGCGATG |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3.1+(Zeo)-FLAG-muGFP was a gift from Daniel Scott (Addgene plasmid # 166726 ; http://n2t.net/addgene:166726 ; RRID:Addgene_166726) |
| 参考文献部分: | A Novel Ultra-Stable, Monomeric Green Fluorescent Protein For Direct Volumetric Imaging of Whole Organs Using CLARITY. Scott DJ, Gunn NJ, Yong KJ, Wimmer VC, Veldhuis NA, Challis LM, Haidar M, Petrou S, Bathgate RAD, Griffin MDW. Sci Rep. 2018 Jan 12;8(1):667. doi: 10.1038/s41598-017-18045-y. 10.1038/s41598-017-18045-y PubMed 29330459 |
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
-FLAG-muGFP质粒图谱.png)
载体简介
载体序列
LOCUS pcDNA3.1+(Zeo)-FLAG-muGFP 5701 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses FLAG-tagged monomeric ultrastable GFP (muGFP) in mammalian
cells.
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1+(Zeo)-FLAG-muGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5701)
AUTHORS Scott DJ, Gunn NJ, Yong KJ, Wimmer VC, Veldhuis NA, Challis LM,
Haidar M, Petrou S, Bathgate RAD, Griffin MDW
TITLE A Novel Ultra-Stable, Monomeric Green Fluorescent Protein For Direct
Volumetric Imaging of Whole Organs Using CLARITY.
JOURNAL Sci Rep. 2018 Jan 12;8(1):667. doi: 10.1038/s41598-017-18045-y.
PUBMED 29330459
REFERENCE 2 (bases 1 to 5701)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5701
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 949..1659
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
ICTTGKLPVPWPTLVTTLTYGVLCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAY
FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLED
VTAAGITHGMDELYK"
primer_bind complement(976..1004)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind complement(1709..1726)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1715..1939
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1985..2413
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2072..2091)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2282..2303
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2422..2442)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2427..2756
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2607..2742
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2669..2688
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 2804..2851
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 2870..3244
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 3374..3495
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3411..3430)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3465..3484
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3544..3560)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3544..3560)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3557..3579)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3568..3584
/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(3592..3622)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3637..3658
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3775..3792)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3946..4534)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4026..4045)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4705..5565)
/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 5328..5347
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5566..5670)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
901 gccaccccat ggatggatta caaagatgat gatgatgtcg acggatccag caaaggagaa
961 gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
1021 aaattttctg tccgtggaga gggtgaaggt gatgctacaa acggaaaact cacccttaaa
1081 tttatttgca ctactggaaa actacctgtt ccgtggccaa cacttgtcac tactctgacc
1141 tatggtgttc tatgcttttc ccgttatccg gatcacatga aacggcatga ctttttcaag
1201 agtgccatgc ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga tgacgggacc
1261 tacaagacgc gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg tatcgagtta
1321 aaaggtattg attttaaaga agatggaaac attctcggac acaaactgga gtacaacttt
1381 aactcacaca atgtatacat cacggcagac aaacaaaaga atggaatcaa agcttacttc
1441 aaaattcgcc acaacgttga agatggttcc gttcaactag cagaccatta tcaacaaaat
1501 actccaattg gcgatggccc tgtcctttta ccagacaacc attacctctc gacacaatct
1561 gttctttcga aagatcccaa cgaaaagcgt gaccacatgg tccttcttga ggatgtaact
1621 gctgctggga ttacacatgg catggatgaa ctctacaaaa ctagctgata actcgagtct
1681 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
1741 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
1801 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
1861 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
1921 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
1981 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
2041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
2221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
2281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
2341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
2401 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
2461 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
2521 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
2581 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
2641 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
2701 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
2761 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
2821 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt
2881 gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac
2941 cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga
3001 cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc
3061 ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac
3121 gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg
3181 ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga
3241 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
3301 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
3361 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
3421 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
3481 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
3541 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
3601 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
3661 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
3721 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
3781 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
3841 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
3901 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
3961 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
4021 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
4081 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
4141 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
4201 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
4261 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
4321 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
4381 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
4441 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
4501 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
4561 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
4621 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
4681 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
4741 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
4801 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
4861 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
4921 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
4981 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
5041 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
5101 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
5161 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
5221 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
5281 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
5341 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
5401 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
5461 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
5521 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
5581 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
5641 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
5701 c
//
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