| 质粒名称: | pBT120a17 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 138510 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV with a BR322 origin |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | Synthetic Biology |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | gIII-xLuxAB |
| 物种来源: | Other |
| 插入大小: | -- |
| 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: | pBT120a17 was a gift from David Liu (Addgene plasmid # 138510 ; http://n2t.net/addgene:138510 ; RRID:Addgene_138510) |
| 参考文献部分: | Phage-assisted evolution of an adenine base editor with improved Cas domain compatibility and activity. Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW, Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA and Liu DR. Nat Biotechnol (2020) 10.1038/s41587-020-0453-z |
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 pBT120a17 6808 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION P1.
ACCESSION .
VERSION .
KEYWORDS pBT120a17
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6808)
AUTHORS Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW,
Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR
TITLE Phage-assisted evolution of an adenine base editor with improved Cas
domain compatibility and activity
JOURNAL Nat Biotechnol (2020)
REFERENCE 2 (bases 1 to 6808)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6808
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 70..89
/label=T7
/note="T7 promoter, forward primer"
promoter 70..88
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1399..2478
/codon_start=1
/gene="luxA"
/product="LuxA luciferase subunit"
/label=LuxA
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
KLFQSDVMPFLKEKQRSLLY"
CDS 2542..3522
/codon_start=1
/gene="luxB"
/product="LuxB luciferase subunit"
/label=LuxB
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="KFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENHF
SDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILGF
SDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAYT
PGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEID
HQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECITA
AKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHTEYT"
misc_RNA complement(3642..3717)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(3744..3774)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3789..3810
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(4022..4882)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQDQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 4645..4664
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4883..4987)
/gene="bla"
/label=AmpR promoter
primer_bind 5055..5073
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
CDS complement(5094..6044)
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFKIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 6092..6314
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
ORIGIN
1 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttgctga ggagacttag
61 ggaccctact aatacgactc actataggga gaaagacctg caggtgcagt aaaggaaaaa
121 aaaaatgaaa aaattattat tcgcaattcc tttagttgtt cctttctatt ctcactccgc
181 tgaaactgtt gaaagttgtt tagcaaaacc ccatacagaa aattcattta ctaacgtctg
241 gaaagacgac aaaactttag atcgttacgc taactatgag ggctgtctgt ggaatgctac
301 aggcgttgta gtttgtactg gtgacgaaac tcagtgttac ggtacatggg ttcctattgg
361 gcttgctatc cctgaaaatg agggtggtgg ctctgagggt ggtggttctg agggtggcgg
421 ttctgagggt ggcggtacta aacctcctga gtacggtgat acacctattc cgggctatac
481 ttatatcaac cctctcgacg gcacttatcc gcctggtact gagcaaaacc ccgctaatcc
541 taatccttct cttgaggagt ctcagcctct taatactttc atgtttcaga ataataggtt
601 ccgaaatagg cagggggcat taactgttta tacgggcact gttactcaag gcactgaccc
661 cgttaaaact tattaccagt acactcctgt atcatcaaaa gccatgtatg acgcttactg
721 gaacggtaaa ttcagagact gcgctttcca ttctggcttt aatgaggatc cattcgtttg
781 tgaatatcaa ggccaatcgt ctgacctgcc tcaacctcct gtcaatgctg gcggcggctc
841 tggtggtggt tctggtggcg gctctgaggg tggtggctct gagggtggcg gttctgaggg
901 tggcggctct gagggaggcg gttccggtgg tggctctggt tccggtgatt ttgattatga
961 aaagatggca aacgctaata agggggctat gaccgaaaat gccgatgaaa acgcgctaca
1021 gtctgacgct aaaggcaaac ttgattctgt cgctactgat tacggtgctg ctatcgatgg
1081 tttcattggt gacgtttccg gccttgctaa tggtaatggt gctactggtg attttgctgg
1141 ctctaattcc caaatggctc aagtcggtga cggtgataat tcacctttaa tgaataattt
1201 ccgtcaatat ttacctagtc ttccgcagag tgtagagtgc cgaccattcg ttttcggtgc
1261 gggcaagccg tacgagttca gcatcgactg cgataagatc aatttgtttc gaggagtttt
1321 cgcattccta ctctacgtgg caacgttcat gtacgtgttc tcaacattcg ccaatatttt
1381 aagaaacaaa gagtcttaat gaaatttgga aactttttgc ttacatacca acctccccaa
1441 ttttcccaaa cagaggtaat gaaacgtttg gttaaattag gtcgcatctc tgaggagtgt
1501 ggttttgata ccgtatggtt actggagcat catttcacgg agtttggttt gcttggtaac
1561 ccttatgtcg ctgctgcata tttacttggc gcgactaaaa aattgaatgt aggaactgcc
1621 gctattgttc ttcccacagc ccatccagta cgccaacttg aagatgtgaa tttattggat
1681 caaatgtcaa aaggacgatt tcggtttggt atttgccgag ggctttacaa caaggacttt
1741 cgcgtattcg gcacagatat gaataacagt cgcgccttag cggaatgctg gtacgggctg
1801 ataaagaatg gcatgacaga gggatatatg gaagctgata atgaacatat caagttccat
1861 aaggtaaaag taaaccccgc ggcgtatagc agaggtggcg caccggttta tgtggtggct
1921 gaatcagctt cgacgactga gtgggctgct caatttggcc taccgatgat attaagttgg
1981 attataaata ctaacgaaaa gaaagcacaa cttgagcttt ataatgaagt ggctcaagaa
2041 tatgggcacg atattcataa tatcgaccat tgcttatcat atataacatc tgtagatcat
2101 gactcaatta aagcgaaaga gatttgccgg aaatttctgg ggcattggta tgattcttat
2161 gtgaatgcta cgactatttt tgatgattca gaccaaacaa gaggttatga tttcaataaa
2221 gggcagtggc gtgactttgt attaaaagga cataaagata ctaatcgccg tattgattac
2281 agttacgaaa tcaatcccgt gggaacgccg caggaatgta ttgacataat tcaaaaagac
2341 attgatgcta caggaatatc aaatatttgt tgtggatttg aagctaatgg aacagtagac
2401 gaaattattg cttccatgaa gctgttccag tctgatgtca tgccatttct taaagaaaaa
2461 caacgttcgc tattatatta tggcggtggc ggtagcggcg gtggcggtag cggcggtggc
2521 ggtagcggcg gtggcggtag caaatttgga ttgttcttcc ttaacttcat caattcaaca
2581 actgttcaag aacagagtat agttcgcatg caggaaataa cggagtatgt tgataagttg
2641 aattttgaac agattttagt gtatgaaaat catttttcag ataatggtgt tgtcggcgct
2701 cctctgactg tttctggttt tctgctcggt ttaacagaga aaattaaaat tggttcatta
2761 aatcacatca ttacaactca tcatcctgtc cgcatagcgg aggaagcttg cttattggat
2821 cagttaagtg aagggagatt tattttaggg tttagtgatt gcgaaaaaaa agatgaaatg
2881 cattttttta atcgcccggt tgaatatcaa cagcaactat ttgaagagtg ttatgaaatc
2941 attaacgatg ctttaacaac aggctattgt aatccagata acgattttta tagcttccct
3001 aaaatatctg taaatcccca tgcttatacg ccaggcggac ctcggaaata tgtaacagca
3061 accagtcatc atattgttga gtgggcggcc aaaaaaggta ttcctctcat ctttaagtgg
3121 gatgattcta atgatgttag atatgaatat gctgaaagat ataaagccgt tgcggataaa
3181 tatgacgttg acctatcaga gatagaccat cagttaatga tattagttaa ctataacgaa
3241 gatagtaata aagctaaaca agagacgcgt gcatttatta gtgattatgt tcttgaaatg
3301 caccctaatg aaaatttcga aaataaactt gaagaaataa ttgcagaaaa cgctgtcgga
3361 aattatacgg agtgtataac tgcggctaag ttggcaattg aaaagtgtgg tgcgaaaagt
3421 gtattgctgt cctttgaacc aatgaatgat ttgatgagcc aaaaaaatgt aatcaatatt
3481 gttgatgata atattaagaa gtaccacacg gaatatacct aaacggagcc aatgtacgca
3541 aaaaccccgc ttcagcgggg ttttttcgcc aaaaaaaacc ccgccctgtc aggggcgggg
3601 ttttttttta tgttgaaaat ctccttctag attaaaaaaa agcaccgact cggtgccact
3661 ttttcaagtt gataacggac tagccttatt ttaacttgct atttctagct ctaaaaccaa
3721 gatgtttgag tgataactcc acacaacata cgagccggaa gcataaagtg taaagcctgg
3781 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
3841 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
3901 ttgcgtattg ggcgcactta attaacggca ctgcgaaaaa accccgccga agcggggttt
3961 tttgcgaaat ataatgaccc tcttgataac ccaagagggc attttttaat gcccatggcg
4021 tttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
4081 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
4141 ccagtgctgc aatgataccg cgtgacccac gctcaccggc tccagattta tcagcaataa
4201 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
4261 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
4321 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
4381 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
4441 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
4501 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
4561 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
4621 gatcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
4681 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
4741 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
4801 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
4861 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
4921 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
4981 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
5041 cattaaccta taaaaatagg cgtatcacga ggcccttagg ccaaataggc cgttcagatc
5101 cttccgtatt tagccagtat gttctctagt gtggttcgtt gtttttgcgt gagccatgag
5161 aacgaaccat tgagatcatg cttactttgc atgtcactca aaaattttgc ctcaaaactg
5221 gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc gttacgtagg
5281 taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat ctggttgttc
5341 tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat caacgtatca
5401 gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt taaatcttta
5461 cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt attttcaagc
5521 attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg agttttcttt
5581 tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt ttcaaaagac
5641 ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata aggcaatatc
5701 tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt tgtccactgg
5761 aaaatcttaa agcctttaac caaaggattc ctgatttcca cagttctcgt catcagctct
5821 ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat catctgagcg
5881 tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc aatcgtgggg
5941 ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa gtcatagcga
6001 ctaatcgcta gttcatttgc tttgaaaaca actaattcag acatacatct caattggtct
6061 aggtgatttt aatcactata ccaattgaga tgggctagtc aatgataatt actagtcctt
6121 ttcctttgag ttgtgggtat ctgtaaattc tgctagacct ttgctggaaa acttgtaaat
6181 tctgctagac cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt ttatattcaa
6241 gtggttataa tttatagaat aaagaaagaa taaaaaaaga taaaaagaat agatcccagc
6301 cctgtgtata actcactact ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg
6361 ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag gcttaagtag caccctcgca
6421 agctcgggca aatcgctgaa tattcctttt gtctccgacc atcaggcacc tgagtcgctg
6481 tctttttcgt gacattcagt tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg
6541 gcactacagg cgccttttat ggattcatgc aaggaaacta cccataatac aagaaaagcc
6601 cgtcacgggc ttctcagggc gttttatggc gggtctgcta tgtggtgcta tctgactttt
6661 tgctgttcag cagttcctgc cctctgattt tccagtctga ccacttcgga ttatcccgtg
6721 acaggtcatt cagactggct aatgcaccca gtaaggcagc ggtatcatca acaggcttac
6781 ccgtcttact gtcaagagga catccggt
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：pCCl-MCS_miniCMV_EGFP
- 下一篇：pBT29
