| 质粒名称: | pCMV-Bxb1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 182142 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 载体出品公司: | Clontech |
| 空载体大小: | 5803 bp |
| 完整质粒大小: | 5801bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | human codon optimized Bxb1 |
| 物种来源: | Synthetic |
| 插入大小: | 1500 bp |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-Bxb1 was a gift from David Liu (Addgene plasmid # 182142 ; http://n2t.net/addgene:182142 ; RRID:Addgene_182142) |
| 参考文献部分: | Programmable deletion, replacement, integration and inversion of large DNA sequences with twin prime editing. Anzalone AV, Gao XD, Podracky CJ, Nelson AT, Koblan LW, Raguram A, Levy JM, Mercer JAM, Liu DR. Nat Biotechnol. 2022 May;40(5):731-740. doi: 10.1038/s41587-021-01133-w. Epub 2021 Dec 9. 10.1038/s41587-021-01133-w PubMed 34887556 |
买家导航
pCMV-Bxb1载体质粒基本信息
pCMV-Bxb1质粒图谱载体图谱和pCMV-Bxb1载体序列质粒序列多克隆位点信息
pCMV-Bxb1质粒载体简介
pCMV-Bxb1质粒序列载体序列
LOCUS pCMV-Bxb1 5801 bp DNA circular SYN 25-AUG-2024
DEFINITION Expression of human codon optimized Bxb1.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5801)
AUTHORS Anzalone AV, Gao XD, Podracky CJ, Nelson AT, Koblan LW, Raguram A,
Levy JM, Mercer JAM, Liu DR
TITLE Programmable deletion, replacement, integration and inversion of
large DNA sequences with twin prime editing.
JOURNAL Nat Biotechnol. 2022 May;40(5):731-740. doi:
10.1038/s41587-021-01133-w. Epub 2021 Dec 9.
PUBMED 34887556
REFERENCE 2 (bases 1 to 5801)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1038/s41587-021-01133-w"; journalName: "Nat Biotechnol"; date:
"2022-05"; volume: "40"; issue: "5"; pages: "731-740"
FEATURES Location/Qualifiers
source 1..5801
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 2..456
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(89..108)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 298..319
/label=F1ori-F
/note="F1 origin, forward primer"
CDS 743..1603
/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 complement(961..980)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
protein_bind 1705..1738
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
rep_origin 1816..2404
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2305..2324
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2558..2575
/label=L4440
/note="L4440 vector, forward primer"
promoter complement(2649..2753)
/gene="bla"
/label=AmpR promoter
primer_bind 2821..2839
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2866..2896
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind 2904..2920
/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)."
primer_bind 2909..2931
/label=M13/pUC Reverse
/note="In lacZ gene"
enhancer 2950..3253
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3254..3457
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 3408..3428
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 3454..3478
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 3573..3591
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 3573..3590
/label=SP6
/note="SP6 promoter, forward primer"
polyA_signal 5543..5677
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5580..5599)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5634..5653
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgcc
541 aggtcttgga ctggtgagaa cggcttgctc ggcagcttcg atgtgtgctg gagggagaat
601 aaaggtctaa gatgtgcgat agagggaagt cgcattgaat tatgtgctgt gtagggatcg
661 ctggtatcaa atatgtgtgc ccacccctgg catgagacaa taaccctgat aaatgcttca
721 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
781 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
841 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
901 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
961 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
1021 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
1081 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
1141 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
1201 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
1261 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
1321 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
1381 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
1441 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
1501 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
1561 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
1621 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
1681 gatccttttt gataatctca tgccataact tcgtataatg tatgctatac gaggttatgg
1741 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
1801 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
1861 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
1921 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
1981 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
2041 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
2101 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
2161 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
2221 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
2281 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
2341 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
2401 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
2461 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
2521 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
2581 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
2641 agcttgcaat tcgcgctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
2701 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
2761 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
2821 aataggcgta tcacgaggcc ctttcactca ttaggcaccc caggctttac actttatgct
2881 tccggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcat
2941 cgtgcaggtc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
3001 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
3061 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
3121 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
3181 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
3241 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
3301 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
3361 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
3421 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
3481 ctggaggcgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
3541 ccggactcta gcctaggctt ttgcaaaaag ctatttaggt gacactatag aaggtacgcc
3601 tgcaggtacc ggtccggaat tcgcccttat gcgggccctg gttgtgatta gactgtccag
3661 agtgaccgac gccaccacct cacctgagag acagttggag agctgccagc agctgtgcgc
3721 ccagcgcggc tgggacgtgg tgggcgtggc cgaggacctg gacgtgtccg gcgccgtgga
3781 cccttttgac cggaagcgga gacctaatct ggccagatgg ctggccttcg aggaacagcc
3841 tttcgatgtg atcgtggcct acagagtgga ccggctgacc agaagcatcc ggcacctgca
3901 acagctggtg cactgggccg aggatcacaa aaagctggtg gtctctgcta cagaggccca
3961 cttcgacaca accacacctt tcgccgctgt ggtgatcgcc ctgatgggga ccgtggccca
4021 gatggaactc gaggccatca aggaacggaa cagatctgcc gcccacttta acatcagagc
4081 cggcaagtac agaggaagcc tgccaccttg gggctacctg cctaccagag ttgacggcga
4141 gtggagactg gtccccgacc ccgtgcagag agagcggatc ctggaggtgt accacagagt
4201 ggtcgacaac cacgagcccc tgcatctggt ggctcatgat ctgaatagaa ggggcgtgct
4261 gagccctaag gactacttcg ctcagctgca gggcagagaa ccccagggca gagagtggag
4321 cgccaccgcc ctgaagagaa gcatgatcag cgaggccatg ctgggatacg ccaccctgaa
4381 cggcaaaacc gtgcgggacg atgatggagc ccctctggtg cgggcagagc ctatcctaac
4441 ccgcgagcag ctggaagctc tgagagccga gctggtcaag acatctagag ccaaacctgc
4501 tgtgtccacc cctagcctgc tgctgcgggt gctgttctgt gccgtgtgcg gtgaacctgc
4561 ttataagttc gccggcggcg gcagaaagca ccccagatac cggtgcagat ctatgggctt
4621 ccccaagcac tgtggcaacg gcaccgtggc catggccgag tgggatgctt tttgcgagga
4681 acaagtgctg gacctgctgg gcgatgccga aagactagag aaggtgtggg tggctggcag
4741 cgacagcgcc gtggaactgg ctgaagtgaa cgccgagctg gtggacctca ccagcctcat
4801 cggaagcccg gcctaccggg ccggctctcc acagagagaa gccctggatg cccgtatcgc
4861 cgccctggcc gctaggcagg aggaactgga aggcctggaa gccagaccaa gcggctggga
4921 gtggagagag acaggacagc ggttcggaga ttggtggcgg gaacaggata cagccgcaaa
4981 gaacacctgg ctgagaagca tgaacgtgcg gctgacattc gacgtgagag gcggactgac
5041 cagaacaatc gacttcggcg acctgcagga gtacgagcaa cacctgagac tgggcagcgt
5101 ggtcgagcgg ctgcacacag gcatgagcta gaagggcgaa ttcccgggtc gacgagctca
5161 ctagcttggg atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact
5221 acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa
5281 actagctgca tatgcttgct gcttgagagt tttgcttact gagtatgatt tatgaaaata
5341 ttatacacag gagctagtga ttctaattgt ttgtgtattt tagattcaca gtcccaaggc
5401 tcatttcagg cccctcagtc ctcacagtct gttcatgatc ataatcagcc ataccacatt
5461 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
5521 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
5581 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
5641 gtccaaactc atcaatgtat cttatcatgt ctggatcgat cctgcattaa tgaatcggcc
5701 aacgcgcggg gagaggcggt ttgcgtattg gctggcgtaa tagcgaagag gcccgcaccg
5761 atcgcccttc ccaacagttg cgcagcctga atggcgaatg g
//
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