| 质粒名称: | pDY0423 Cargo EGFP with AttP Bxb1 site for in-frame fusion (Parent minicircle) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 179116 |
| 存储实验室: | Omar Abudayyeh, Jonathan Gootenberg |
|
载体骨架基本信息 |
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| 载体名称: | pMC.BESPX |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 4308 bp |
| 载体修饰: | -- |
| 载体类型: | CRISPR |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素，50μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB稳定 |
| 拷贝数: | -- |
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插入基因信息 |
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| 插入基因名称: | Cargo EGFP with AttP Bxb1 site for in-frame fusion |
|
文献引用方法 |
|
| 材料和方法部分: | pDY0423 Cargo EGFP with AttP Bxb1 site for in-frame fusion (Parent minicircle) was a gift from Omar Abudayyeh & Jonathan Gootenberg (Addgene plasmid # 179116 ; http://n2t.net/addgene:179116 ; RRID:Addgene_179116) |
| 参考文献部分: | Drag-and-drop genome insertion of large sequences without double-strand DNA cleavage using CRISPR-directed integrases. Yarnall MTN, Ioannidi EI, Schmitt-Ulms C, Krajeski RN, Lim J, Villiger L, Zhou W, Jiang K, Garushyants SK, Roberts N, Zhang L, Vakulskas CA, Walker JA 2nd, Kadina AP, Zepeda AE, Holden K, Ma H, Xie J, Gao G, Foquet L, Bial G, Donnelly SK, Miyata Y, Radiloff DR, Henderson JM, Ujita A, Abudayyeh OO, Gootenberg JS. Nat Biotechnol. 2022 Nov 24. doi: 10.1038/s41587-022-01527-4. 10.1038/s41587-022-01527-4 PubMed 36424489 |
pDY0423 Cargo EGFP with AttP Bxb1 site for in-frame fusion (
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
质粒图谱.png)
载体简介
载体序列
LOCUS pDY0423 Cargo EGFP with AttP Bxb1 site for in-frame fusion (Parent minicircle) 4308 bp ds-DNA circular SYN 11-JAN-2022
DEFINITION Cargo EGFP with AttP Bxb1 site for in-frame fusion (Parent
minicircle).
ACCESSION .
VERSION .
KEYWORDS pDY0423 Cargo EGFP with AttP Bxb1 site for in-frame fusion (Parent minicircle)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4308)
AUTHORS Ioannidi EI, Yarnall MTN, Schmitt-Ulms C, Krajeski RN, Lim J,
Villiger L, Zhou W, Jiang K, Roberts N, Zhang L, Vakulskas CA,
Walker II JA, Kadina AP, Zepeda AE, Holden K, Gootenberg JS,
Abudayyeh OO
TITLE Drag-and-drop genome insertion without DNA cleavage with
CRISPR-directed integrases
JOURNAL bioRxiv 2021.11.01.466786
REFERENCE 2 (bases 1 to 4308)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 11, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4308
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(328..345)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 498..584
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 676..703
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
rep_origin 837..1425
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1326..1345
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1579..1596
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 1611..1751
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(1677..1696)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1837..1859
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2538..2555)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
protein_bind 2576..2609
/label=attB
/bound_moiety="phage phi-C31 integrase"
/note="minimal attB site for the phi-C31 integrase (Groth
et al., 2000)"
CDS 2619..3284
/codon_start=1
/product="green fluorescent protein 2 from Pontellina
plumata, also known as ppluGFP2 (Shagin et al., 2004)"
/label=CopGFP
/translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
"
CDS complement(3478..4272)
/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
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3589..3608)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 4199..4218
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
ORIGIN
1 tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg
61 cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa
121 aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc
181 gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg
241 tttctgcgga ctggctttct acgcgctcga gggggggcca aacggtctcc agcttggctg
301 ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
361 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
421 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
481 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
541 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
601 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
661 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
721 ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaccaa aatcccttaa
781 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
841 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
901 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
961 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
1021 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
1081 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
1141 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
1201 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
1261 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
1321 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
1381 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
1441 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
1501 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
1561 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
1621 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
1681 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
1741 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
1801 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
1861 tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc
1921 ggcatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact
1981 ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg gctacatcat
2041 tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa
2101 atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag
2161 gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc
2221 ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc
2281 aaacatgctg tgcgacgctg gcgattcttc tctcatccgc caaaacagcc aagctggaga
2341 ccgtttgaca ttaccctgtt atccctagat acattaccct gttatcccag atgacatacc
2401 ctgttatccc tagatgacat taccctgtta tcccagatga cattaccctg ttatccctag
2461 atacattacc ctgttatccc agatgacata ccctgttatc cctagatgac attaccctgt
2521 tatcccagat aaactcaatg atgatgatga tgatggtcga gactcagcgg ccgcggtgcc
2581 agggcgtgcc cttgggctcc ccgggcgcgg ccgccaccat gcccgccatg aagatcgagt
2641 gccgcatcac cggcaccctg aacggcgtgg agttcgagct ggtgggcggc ggagagggca
2701 cccccgagca gggccgcatg accaacaaga tgaagagcac caaaggcgcc ctgaccttca
2761 gcccctacct gctgagccac gtgatgggct acggcttcta ccacttcggc acctacccca
2821 gcggctacga gaaccccttc ctgcacgcca tcaacaacgg cggctacacc aacacccgca
2881 tcgagaagta cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc tacgaggccg
2941 gccgcgtgat cggcgacttc aaggtggtgg gcaccggctt ccccgaggac agcgtgatct
3001 tcaccgacaa gatcatccgc agcaacgcca ccgtggagca cctgcacccc atgggcgata
3061 acgtgctggt gggcagcttc gcccgcacct tcagcctgcg cgacggcggc tactacagct
3121 tcgtggtgga cagccacatg cacttcaaga gcgccatcca ccccagcatc ctgcagaacg
3181 ggggccccat gttcgccttc cgccgcgtgg aggagctgca cagcaacacc gagctgggca
3241 tcgtggagta ccagcacgcc ttcaagaccc ccatcgcctt cgccagatct cgagctcgag
3301 gtggtttgtc tggtcaacca ccgcggtctc agtggtgtac ggtacaaacc caccccaact
3361 ggggtaacct ttgagttctc tcagttgggg gtaatcagca tcatgatgtg gtaccacatc
3421 atgatgctga ttataagaat gcggccgcca cactctagtg gatctcgagt taataattca
3481 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
3541 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
3601 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
3661 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
3721 gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag
3781 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
3841 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
3901 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
3961 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
4021 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
4081 tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
4141 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
4201 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
4261 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagagct
//
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