| 质粒名称: | pcDNA-NBid-PhoCl2c-CBid |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 164051 |
| 存储实验室: | Robert Campbell |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1+ |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5350 bp |
| 完整质粒大小: | 6709 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | NBid-PhoCl2c-CBid |
| 物种来源: | Synthetic |
| 插入大小: | 1359 bp |
| 启动子 | CMV promotor |
| 标签/融合蛋白: | kozak sequence（插入件上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NheI (未破坏) |
| 3' 克隆位点: | XhoI (未破坏) |
| 5' 测序引物 | CGCAAATGGGCGGTAGGCGTG |
| 3' 测序引物 | CCTCGACTGTGCCTTCTA |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA-NBid-PhoCl2c-CBid was a gift from Robert Campbell (Addgene plasmid # 164051 ; http://n2t.net/addgene:164051 ; RRID:Addgene_164051) |
| 参考文献部分: | Photocleavable Proteins that Undergo Fast and Efficient Dissociation. Lu X, Wen Y, Zhang S, Zhang W, Chen Y, Shen Y, Lemieux MJ, Campbell RE. Chem. Sci., 2021, Advance Article 10.1039/D1SC01059J |
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 pcDNA-NBid-PhoCl2c-CBid 6730 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Optogenetic manipulation of cell apoptosis using PhoCl2c.
ACCESSION .
VERSION .
KEYWORDS pcDNA-NBid-PhoCl2c-CBid
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6730)
AUTHORS Lu X, Wen Y, Zhang S, Zhang W, Chen Y, Shen Y, Lemieux MJ, Campbell
RE
TITLE Improved Photocleavable Proteins with Faster and More Efficient
Dissociation
JOURNAL bioRxiv, December 10, 2020.
REFERENCE 2 (bases 1 to 6730)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6730
/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"
regulatory 901..910
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind complement(2321..2338)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2327..2551
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2597..3025
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2684..2703)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2894..2915
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3034..3054)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3039..3368
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3219..3354
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3281..3300
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3435..4229
/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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3489..3508)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4099..4118
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4403..4524
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4440..4459)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4494..4513
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4573..4589)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4573..4589)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4586..4608)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4597..4613
/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(4621..4651)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4666..4687
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4804..4821)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4975..5563)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5055..5074)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5734..6594)
/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 6357..6376
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6595..6699)
/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 gctggctagc
901 gccaccatgg attgtgaggt caataacggt tcatctctcc gagacgaatg cataacgaac
961 ttgctcgttt tcggctttct gcaatcctgc agcgataatt ctttcagaag agaacttgac
1021 gctcttggac atgaactccc agtactcgct ccacagtggg aaggctacga tgagggatcc
1081 gtgatccctg actacttcaa gcagagcttc cccgagggct acagctggga gcgcagcatg
1141 acctacgagg acggcggcat ctgcatcgcc accaacgaca tcacaatgga gggggacagc
1201 ttcatcaaca agatccactt ccagggcacg aacttccccc ccaacggccc cgtgatgcag
1261 aagaggaccg tgggctggga ggccagcacc gagaagatgt acgagcgcga cggcgtgctg
1321 aagggcgacg tgaagatgaa gctgctgctg aagggcggcg gccactatcg cggcgactac
1381 cgcaccacct acaaggtcaa gcagaagccc gtaaagctgc ccgactgcca cttcgtggac
1441 caccgcatcg agatcctgag ccacgacaag gactacaaca aggtgaagct gtacgagcac
1501 gccgtggcca agacttccac cgacagcatg gacgagctgt acaagggtgg cagcggtggc
1561 atggtgagca agggcgagga gaccattaca agcgtgatca agcctgacat gaagaacaag
1621 ctgcgcatgg agggcaacgt gaacggccac gccttcgtga tcgagggcga gggcagcggc
1681 aagcccttcg agggcatcca gacgattgat ttggaggtga aggagggcgc cccgctgccc
1741 ttcgcctacg acatcctgac caccgccttc cactacggca accgcgtgtt caccaagtac
1801 ccacggggta cccttcaaac cgatggaaac cgatcatctc attcaaggtt ggggcgaatt
1861 gaagcagata gtgagagtca ggaggacata atacgcaata tagctcgaca ccttgcacag
1921 gtcggtgata gcatggaccg ctctatcccc ccaggtttgg ttaacggatt ggccctgcaa
1981 ctgcgaaaca cttcaaggag tgaagaagat agaaatcgag acctggcgac cgcgcttgaa
2041 cagcttcttc aagcctatcc cagagatatg gagaaggaaa aaacaatgct cgtgctcgca
2101 ctgctgctgg caaagaaagt agcctctaac acaccatccc tcttgagaga cgtcttccat
2161 accacggtaa atttcataaa ccagaacctt aggacgtatg tgcggagtct tgctcgcaat
2221 ggtatgactg agtttcttca acccgtgccc ctcaatgcgt ggttgctcag gttgaacaag
2281 taactcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
2341 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
2401 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
2461 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
2521 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
2581 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
2641 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
2701 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
2761 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
2821 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
2881 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
2941 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
3001 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
3061 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
3121 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
3181 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
3241 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
3301 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
3361 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
3421 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
3481 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
3541 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
3601 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
3661 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
3721 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
3781 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
3841 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
3901 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
3961 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
4021 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
4081 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
4141 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
4201 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
4261 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
4321 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
4381 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
4441 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
4501 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
4561 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
4621 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
4681 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
4741 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
4801 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
4861 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
4921 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
4981 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
5041 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
5101 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
5161 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
5221 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
5281 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
5341 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
5401 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
5461 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
5521 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
5581 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
5641 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
5701 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
5761 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
5821 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
5881 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
5941 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
6001 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
6061 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
6121 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
6181 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
6241 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
6301 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
6361 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
6421 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
6481 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
6541 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
6601 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
6661 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
6721 acctgacgtc
//
