| 质粒名称: | pBT29 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 138521 |
| 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: | pBT29 was a gift from David Liu (Addgene plasmid # 138521 ; http://n2t.net/addgene:138521 ; RRID:Addgene_138521) |
| 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 pBT29 5662 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION phage cloning.
ACCESSION .
VERSION .
KEYWORDS pBT29
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5662)
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 5662)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5662
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 23..1255
/codon_start=1
/gene="II"
/product="pII"
/label=M13 gene II
/note="replication"
/translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGMNLSAR
TVEYHVDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
FGTTDLGLCSEALLLNFANSLPCLYDLLDVNATTISRIDAAFSARAPNENIAKQVIDHL
RNVSNGQTKSTRSQNWESTVTWNETSRHRTLVAYLKHVELQHQIQQLSSKPSAKMTSYQ
KEQLKVLSNPDLLEFASGLVRFEARIKTRYLKSFGLPLNLFDAIRFASDYNSQGKDLIF
DLWSFSFSELFKAFEGNSMNIYDDSAVLAAIQSKHFTITPSGKTSFAKASRYFGFYRRL
VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
DWYNEPVLKIA"
CDS 1267..1530
/codon_start=1
/gene="V"
/product="pV"
/label=M13 gene V
/note="replication"
/translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQFCYVDLGNEYPVLV
KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
CDS 1532..1633
/codon_start=1
/gene="VII"
/product="pVII"
/label=M13 gene VII
/note="minor coat protein"
/translation="MEQVADFDIIYQAMIQISVVLCFALGIIAGGQR"
CDS 1630..1728
/codon_start=1
/gene="IX"
/product="pIX"
/label=M13 gene IX
/note="minor coat protein"
/translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
CDS 1725..1946
/codon_start=1
/gene="VIII"
/product="pVIII"
/label=M13 gene VIII
/note="major coat protein"
/translation="MKKSLVLKASVAVATLVPMLSFAAEGDDPAKAAFDSLQASATEYI
GYAWAMVVVIVGATIGIKLFKKFTSKAS"
terminator 1953..1992
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
protein_bind 2003..2021
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2028..2046
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
RBS 2064..2075
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 2082..2759
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
terminator 2777..2848
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2864..2891
/label=T7Te terminator
/note="phage T7 early transcription terminator"
CDS complement(2990..3805)
/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 3713..3732
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 3797..3816
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS complement(3873..4823)
/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 4871..5093
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
ORIGIN
1 cttattatca accggggtac atatgattga catgctagtt ttacggttac cgttcatcga
61 ttctcttgtt tgctccagac tctcaggcaa tgacctgata gcctttgtag atctctcaaa
121 aatagctacc ctctccggca tgaatttatc agctagaacg gttgaatatc atgttgatgg
181 tgatttgact gtctccggcc tttcccaccc ttttgaatct ttacctacac attactcagg
241 cattgcattt aaaatatatg agggttctaa aaatttttat ccttgcgttg aaataaaggc
301 ttctcccgca aaagtattac aggggcataa tgtttttggt acaaccgatt taggtttatg
361 ctctgaggct ttattgctta attttgctaa ttctttgcct tgcctgtatg atttattgga
421 tgttaacgct actactatta gtagaattga tgccgccttt tcagcccgcg ccccaaatga
481 aaatatagct aaacaggtta ttgaccattt gcgaaatgta tctaatggtc aaactaaatc
541 tactcgttcg cagaattggg aatcaactgt tacatggaat gaaacttcca gacaccgtac
601 tttagttgca tatttaaaac atgttgagct acagcaccag attcagcaat taagctctaa
661 gccatccgca aaaatgacct cttatcaaaa ggaacaatta aaggtactct ctaatcctga
721 cctgttggag tttgcttccg gtctggttcg ctttgaggct cgaattaaaa cgagatattt
781 gaagtctttc gggcttcctc ttaatctttt tgatgcaatc cgctttgctt ctgactataa
841 tagtcagggt aaagacctga tttttgattt atggtcattc tcgttttctg aactgtttaa
901 agcatttgag gggaattcaa tgaatattta tgacgattcc gcagtattgg ccgctatcca
961 gtctaaacat tttactatta ccccctctgg caaaacttct tttgcaaaag cctctcgcta
1021 ttttggtttt tatcgtcgtc tggtaaacga gggttatgat agtgttgctc ttactatgcc
1081 tcgtaattcc ttttggcgtt atgtatctgc attagttgaa tgtggtattc ctaaatctca
1141 actgatgaat ctttctacct gtaataatgt tgttccgtta gttcgtttta ttaacgtaga
1201 tttttcttcc caacgtcctg actggtataa tgagccagtt cttaaaatcg cataaggtaa
1261 ttcacaatga ttaaagttga aattaaacca tctcaagccc aatttactac tcgttctggt
1321 gtttctcgtc agggcaagcc ttattcactg aatgagcagt tttgttacgt tgatttgggt
1381 aatgaatatc cggttcttgt caagattact cttgatgaag gtcagccagc ctatgcgccg
1441 ggtctgtaca ccgttcatct gtcctctttc aaagttggtc agttcggttc ccttatgatt
1501 gaccgtctgc gcctcgttcc ggctaagtaa catggagcag gtcgcggatt tcgacataat
1561 ttatcaggcg atgatacaaa tctccgttgt actctgtttc gcgcttggta taatagctgg
1621 gggtcaaaga tgagtgtttt agtgtattct ttcgcctctt tcgttttagg ttggtgcctc
1681 cgtagtggca ttacgtattt tacccgttta atggaaactt cctcatgaaa aagtctttag
1741 tcctcaaagc ctctgtagcc gttgctaccc tcgttccgat gctgtctttc gctgctgagg
1801 gtgacgatcc cgcaaaagcg gcctttgact ccctgcaagc ctcagcgacc gaatatatcg
1861 gttatgcgtg ggcgatggtt gttgtcattg tcggcgcaac tatcggtatc aagctgttta
1921 agaaattcac ctcgaaagca agttgataaa ctgatacaat taaaggctcc ttttggagcc
1981 tttttttttg gagtagaaga gctccctatc agtgatagag attgacatcc ctatcagtga
2041 tagagatact gagcacgaat ctgaaagagg agaaaggatc tatggcgagt agcgaagacg
2101 ttatcaaaga gttcatgcgt ttcaaagttc gtatggaagg ttccgttaac ggtcacgagt
2161 tcgaaatcga aggtgaaggt gaaggtcgtc cgtacgaagg tacccagacc gctaaactga
2221 aagttaccaa aggtggtccg ctgccgttcg cttgggacat cctgtccccg cagttccagt
2281 acggttccaa agcttacgtt aaacacccgg ctgacatccc ggactacctg aaactgtcct
2341 tcccggaagg tttcaaatgg gaacgtgtta tgaacttcga agacggtggt gttgttaccg
2401 ttacccagga ctcctccctg caagacggtg agttcatcta caaagttaaa ctgcgtggta
2461 ccaacttccc gtccgacggt ccggttatgc agaaaaaaac catgggttgg gaagcttcca
2521 ccgaacgtat gtacccggaa gacggtgctc tgaaaggtga aatcaaaatg cgtctgaaac
2581 tgaaagacgg tggtcactac gacgctgaag ttaaaaccac ctacatggct aaaaaaccgg
2641 ttcagctgcc gggtgcttac aaaaccgaca tcaaactgga catcacctcc cacaacgaag
2701 actacaccat cgttgaacag tacgaacgtg ctgaaggtcg tcactccacc ggtgcttaat
2761 aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
2821 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
2881 gcctttctgc gtttatacag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
2941 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
3001 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
3061 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
3121 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
3181 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
3241 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
3301 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
3361 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
3421 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
3481 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
3541 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
3601 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
3661 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
3721 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
3781 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
3841 gttttattgt tcatgaaggc caaataggcc gttcagatcc ttccgtattt agccagtatg
3901 ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt gagatcatgc
3961 ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt
4021 aaagcatcgt gtagtgtttt tcttagtccg ttacgtaggt aggaatctga tgtaatggtt
4081 gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt tacgagatcc
4141 atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca
4201 accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt caaaacccat
4261 tggttaagcc ttttaaactc atggtagtta ttttcaagca ttaacatgaa cttaaattca
4321 tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc ttttaataac
4381 cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc cagattatat
4441 tttatgaatt tttttaactg gaaaagataa ggcaatatct cttcactaaa aactaattct
4501 aatttttcgc ttgagaactt ggcatagttt gtccactgga aaatctcaaa gcctttaacc
4561 aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt agctaataca
4621 ccataagcat tttccctact gatgttcatc atctgagcgt attggttata agtgaacgat
4681 accgtccgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc cacacagcat
4741 aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag ttcatttgct
4801 ttgaaaacaa ctaattcaga catacatctc aattggtcta ggtgatttta atcactatac
4861 caattgagat gggctagtca atgataatta ctagtccttt tcctttgagt tgtgggtatc
4921 tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat
4981 tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat ttatagaata
5041 aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa ctcactactt
5101 tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac
5161 agaccttaaa accctaaagg cttaagtagc accctcgcaa gctcgggcaa atcgctgaat
5221 attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg acattcagtt
5281 cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc gccttttatg
5341 gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct tctcagggcg
5401 ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc
5461 ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta
5521 atgcacccag taaggcagcg gtatcatcaa caggcttacc cgtcttactg tcaagaggac
5581 atccggtcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttgctgagga
5641 gacttaggga ccctgctctt ct
//
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