| 质粒名称: | pBT114 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 138523 |
| 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: | 卡那霉素50 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: | phage genes |
| 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: | pBT114 was a gift from David Liu (Addgene plasmid # 138523 ; http://n2t.net/addgene:138523 ; RRID:Addgene_138523) |
| 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 pBT114 6881 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION phage cloning.
ACCESSION .
VERSION .
KEYWORDS pBT114
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6881)
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 6881)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6881
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 983..1437
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1070..1089)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1279..1300
/label=F1ori-F
/note="F1 origin, forward primer"
protein_bind 1489..1507
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1514..1532
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
RBS 1550..1561
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 1568..2245
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
terminator 2263..2334
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2350..2377
/label=T7Te terminator
/note="phage T7 early transcription terminator"
CDS complement(2476..3291)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 3199..3218
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 3283..3302
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS complement(3359..4309)
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 4357..4579
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
CDS 5232..5570
/codon_start=1
/gene="VI"
/product="pVI"
/label=M13 gene VI
/note="minor coat protein"
/translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKTAIFIFDVKQKI
VSYLDWDK"
CDS 5572..6618
/codon_start=1
/gene="I"
/product="pI"
/label=M13 gene I
/note="phage assembly"
/translation="MAVYFVTGKLGSGKTLVSVGKIQDKIVAGCKIATNLDLRLQNLPQ
VGRFAKTPRVLRIPDKPSISDLLAIGRGNDSYDENKNGLLVLDECGTWFNTRSWNDKER
QPIIDWFLHARKLGWDIIFLVQDLSIVDKQARSALAEHVVYCRRLDRITLPFVGTLYSL
ITGSKMPLPKLHVGVVKYGDSQLSPTVERWLYTGKNLYNAYDTKQAFSSNYDSGVYSYL
TPYLSHGRYFKPLNLGQKMKLTKIYLKKFSRVLCLAIGFASAFTYSYITQPKPEVKKVV
SQTYDFDKFTIDSSHRLNLSYRYVFKDSKGKLINSDDLQKQGYSLTYIDLCTVSIKKGN
SNEIVKCN"
CDS 6596..995
/codon_start=1
/gene="IV"
/product="pIV"
/label=M13 gene IV
/note="phage assembly"
/translation="MKLLNVINFVFLMFVSSSSFAQVIEMNNSPLRDFITWYSKQSGES
VIVSPDVKGTVTVYSSDVKPENLRNFFISVLRANNFDMVGSNPSIIQKYNPNNQDYIDE
LPSPDNQEYDDNSAPSGGFFVPQNDNVTQTFKINNVRAKDLIRVVELFVKSNTSKSSNV
LSIDGSNLLVVSAPKDILDNLSQFLSTVDLPTDQVLIEGLIFEVQQGDALDFSFAAGSQ
RGTVAGGVNTDRLTSVLSSAGGSFGIFNGDVLGLSVRALKTNSHSKILSVPRILTLSGQ
KGSISIGQNVPFITGRVTGESANVNNPFQTIERQNVGISMSVFPVAMAGGNIVLDITSK
ADSLSSSTQASDVITNQRSIATTVNLRDGQTLLLGGLTDYKNTSQDSGVPFLSKIPLIG
LLFSSRSDSNEESTLYVLVKATIVRAL"
ORIGIN
1 caaacaatca ggattatatt gatgaattgc catcacctga taatcaggaa tatgatgata
61 attccgctcc ttctggtggt ttctttgtcc cgcaaaatga taatgttact caaactttta
121 aaattaataa cgttcgggca aaggatttaa tacgagttgt cgaattgttt gtaaagtcta
181 atacttctaa atcctcaaat gtattatcta ttgacggatc taatctatta gttgttagtg
241 ctcctaaaga tattttagat aacctttctc aattcctttc aactgttgat ttgccaactg
301 accaggtatt gattgagggt ttgatatttg aggttcagca aggtgatgct ttagattttt
361 catttgctgc tggctctcag cgtggcactg ttgcaggcgg agttaatact gaccgcctca
421 cctctgtttt atcttctgct ggtggttcgt tcggtatttt taatggcgat gttttagggc
481 tatcagttcg cgcattaaag actaatagcc attcaaaaat attgtctgtg ccacgtattc
541 ttacgctttc aggtcagaag ggttctatct ctattggcca gaatgtccct tttattactg
601 gtcgtgtaac tggtgaatct gccaatgtaa ataatccatt tcagacgatt gagcgtcaaa
661 atgtaggtat ttccatgagc gtttttcctg ttgcaatggc tggcggtaat attgttctgg
721 atattaccag caaggccgat agtttgagtt cttctactca ggcaagtgat gttattacta
781 accaaagaag tattgctaca acggttaatt tgcgtgatgg acagactctt ctactcggtg
841 gcctcactga ttataaaaac acttctcagg attctggcgt accgttcctg tctaaaatcc
901 ctttaatcgg cctcctgttt agctcccgct ctgattctaa cgaggagagc acgttatacg
961 tgctcgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
1021 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
1081 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagcgct aaatcggggg
1141 cccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
1201 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
1261 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
1321 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
1381 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa
1441 tatttgctta tacaatcttc ctgtttttgg ggcttttctt agaagagctc cctatcagtg
1501 atagagattg acatccctat cagtgataga gatactgagc acgaatctga aagaggagaa
1561 aggatctatg gcgagtagcg aagacgttat caaagagttc atgcgtttca aagttcgtat
1621 ggaaggttcc gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta
1681 cgaaggtacc cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg
1741 ggacatcctg tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga
1801 catcccggac tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa
1861 cttcgaagac ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt
1921 catctacaaa gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa
1981 aaaaaccatg ggttgggaag cttccaccga acgtatgtac ccggaagacg gtgctctgaa
2041 aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa
2101 aaccacctac atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa
2161 actggacatc acctcccaca acgaagacta caccatcgtt gaacagtacg aacgtgctga
2221 aggtcgtcac tccaccggtg cttaataagg atctccaggc atcaaataaa acgaaaggct
2281 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
2341 agtcacactg gctcaccttc gggtgggcct ttctgcgttt atacaggtgg cacttttcgg
2401 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
2461 ctcataaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
2521 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
2581 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
2641 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
2701 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
2761 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
2821 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
2881 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
2941 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
3001 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
3061 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
3121 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
3181 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
3241 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
3301 tgtattactg tttatgtaag cagacagttt tattgttcat gaaggccaaa taggccgttc
3361 agatccttcc gtatttagcc agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc
3421 atgagaacga accattgaga tcatgcttac tttgcatgtc actcaaaaat tttgcctcaa
3481 aactggtgag ctgaattttt gcagttaaag catcgtgtag tgtttttctt agtccgttac
3541 gtaggtagga atctgatgta atggttgttg gtattttgtc accattcatt tttatctggt
3601 tgttctcaag ttcggttacg agatccattt gtctatctag ttcaacttgg aaaatcaacg
3661 tatcagtcgg gcggcctcgc ttatcaacca ccaatttcat attgctgtaa gtgtttaaat
3721 ctttacttat tggtttcaaa acccattggt taagcctttt aaactcatgg tagttatttt
3781 caagcattaa catgaactta aattcatcaa ggctaatctc tatatttgcc ttgtgagttt
3841 tcttttgtgt tagttctttt aataaccact cataaatcct catagagtat ttgttttcaa
3901 aagacttaac atgttccaga ttatatttta tgaatttttt taactggaaa agataaggca
3961 atatctcttc actaaaaact aattctaatt tttcgcttga gaacttggca tagtttgtcc
4021 actggaaaat ctcaaagcct ttaaccaaag gattcctgat ttccacagtt ctcgtcatca
4081 gctctctggt tgctttagct aatacaccat aagcattttc cctactgatg ttcatcatct
4141 gagcgtattg gttataagtg aacgataccg tccgttcttt ccttgtaggg ttttcaatcg
4201 tggggttgag tagtgccaca cagcataaaa ttagcttggt ttcatgctcc gttaagtcat
4261 agcgactaat cgctagttca tttgctttga aaacaactaa ttcagacata catctcaatt
4321 ggtctaggtg attttaatca ctataccaat tgagatgggc tagtcaatga taattactag
4381 tccttttcct ttgagttgtg ggtatctgta aattctgcta gacctttgct ggaaaacttg
4441 taaattctgc tagaccctct gtaaattccg ctagaccttt gtgtgttttt tttgtttata
4501 ttcaagtggt tataatttat agaataaaga aagaataaaa aaagataaaa agaatagatc
4561 ccagccctgt gtataactca ctactttagt cagttccgca gtattacaaa aggatgtcgc
4621 aaacgctgtt tgctcctcta caaaacagac cttaaaaccc taaaggctta agtagcaccc
4681 tcgcaagctc gggcaaatcg ctgaatattc cttttgtctc cgaccatcag gcacctgagt
4741 cgctgtcttt ttcgtgacat tcagttcgct gcgctcacgg ctctggcagt gaatgggggt
4801 aaatggcact acaggcgcct tttatggatt catgcaagga aactacccat aatacaagaa
4861 aagcccgtca cgggcttctc agggcgtttt atggcgggtc tgctatgtgg tgctatctga
4921 ctttttgctg ttcagcagtt cctgccctct gattttccag tctgaccact tcggattatc
4981 ccgtgacagg tcattcagac tggctaatgc acccagtaag gcagcggtat catcaacagg
5041 cttacccgtc ttactgtcaa gaggacatcc ggtcaaataa aacgaaaggc tcagtcgaaa
5101 gactgggcct ttcgttttgc tgaggagact tagggaccct gctcttctgg ctaatggaga
5161 ttttcaacat gggctagcac agccctaggt attatgctag cgtggtgtct gcgtaataag
5221 gagtcttaat catgccagtt cttttgggta ttccgttatt attgcgtttc ctcggtttcc
5281 ttctggtaac tttgttcggc tatctgctta cttttctcaa aaagggcttc ggtaagatag
5341 ctattgctat ttcattgttt cttgctctta ttattgggct taactcaatt cttgtgggtt
5401 atctctctga tattagtgct caattaccct ctgactttgt tcagggtgtt cagttaattc
5461 tcccgtctaa tgcgcttccc tgtttttatg ttattctctc tgtaaagact gctattttca
5521 tttttgacgt taaacaaaaa atcgtttctt atttggattg ggacaaataa tatggctgtt
5581 tattttgtaa ctggcaaatt aggctctgga aagacgctcg ttagcgttgg taagattcag
5641 gataaaattg tagctgggtg caaaatagca actaatcttg atttaaggct tcaaaacctc
5701 ccgcaagtcg ggaggttcgc taaaacgcct cgcgttctta gaataccgga taagccttct
5761 atatctgatt tgcttgctat tgggcgcggt aatgattcct acgatgaaaa taaaaacgga
5821 ttgcttgttc tcgatgagtg cggtacttgg tttaataccc gttcttggaa tgataaggaa
5881 agacagccga ttattgattg gtttctacat gctcgtaaat taggatggga tattattttt
5941 cttgttcagg acttatctat tgttgataaa caggcgcgtt ctgcattagc tgaacatgtt
6001 gtttattgtc gtcgtctgga cagaattact ttaccttttg tcggtacttt atattctctt
6061 attactggct cgaaaatgcc tctgcctaaa ttacatgttg gcgttgttaa atatggcgat
6121 tctcaattaa gccctactgt tgagcgttgg ctttatactg gtaagaattt gtataacgca
6181 tacgatacta aacaggcttt ttctagtaat tatgattccg gtgtttattc ttatttaacg
6241 ccttatttat cacacggtcg gtatttcaaa ccattaaatt taggtcagaa gatgaaatta
6301 actaaaatat atttgaaaaa gttttctcgc gttctttgtc ttgcgattgg atttgcatca
6361 gcatttacat atagttatat aacccaacct aagccggagg ttaaaaaggt agtctctcag
6421 acctatgatt ttgataaatt cactattgac tcttctcatc gtcttaatct aagctatcgc
6481 tatgttttca aggattctaa gggaaaatta attaatagcg acgatttaca gaagcaaggt
6541 tattcactta catatattga tttatgtact gtttccatta aaaaaggtaa ttcaaatgaa
6601 attgttaaat gtaattaatt ttgttttctt gatgtttgtt tcatcatctt cttttgctca
6661 ggtaattgaa atgaataatt cgcctctgcg cgattttata acttggtatt caaagcaatc
6721 aggcgaatcc gttattgttt ctcccgatgt aaaaggtact gttactgtat attcatctga
6781 cgttaaacct gaaaatctac gcaatttctt tatttctgtt ttacgtgcaa ataattttga
6841 tatggtaggt tctaaccctt ccattattca gaagtataat c
//
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