| 质粒名称: | pHelper_ShCAST_sgRNA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 127921 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 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: | -- |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素,100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | ShTnsB, ShTnsC, ShTniQ, ShCas12k, sgRNA scaffold for guide cloning (LguI sites) |
| 物种来源: | -- |
| 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: | pHelper_ShCAST_sgRNA was a gift from Feng Zhang (Addgene plasmid # 127921 ; http://n2t.net/addgene:127921 ; RRID:Addgene_127921) |
| 参考文献部分: | RNA-guided DNA insertion with CRISPR-associated transposases. Strecker J, Ladha A, Gardner Z, Schmid-Burgk JL, Makarova KS, Koonin EV, Zhang F. Science. 2019 Jul 5;365(6448):48-53. doi: 10.1126/science.aax9181. Epub 2019 Jun 6. 10.1126/science.aax9181 PubMed 31171706 |
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 pHelper_ShCAST_sgRNA 7712 bp ds-DNA circular SYN 25-MAY-2021
DEFINITION Expresses ShCAST and an empty sgRNA scaffold. New targets can be
added using Golden Gate assembly (LguI sites)..
ACCESSION .
VERSION .
KEYWORDS pHelper_ShCAST_sgRNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7712)
AUTHORS Strecker J, Ladha A, Gardner Z, Schmid-Burgk JL, Makarova KS, Koonin
EV, Zhang F
TITLE RNA-guided DNA insertion with CRISPR-associated transposases.
JOURNAL Science. 2019 Jul 5;365(6448):48-53. doi: 10.1126/science.aax9181.
Epub 2019 Jun 6.
PUBMED 31171706
REFERENCE 2 (bases 1 to 7712)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 25, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7712
/organism="synthetic DNA construct"
/mol_type="other DNA"
RBS 22..33
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
terminator 5451..5522
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
promoter 5537..5570
/label=J23119 promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119)"
terminator 5825..5852
/label=T7Te terminator
/note="phage T7 early transcription terminator"
promoter 5879..5983
/gene="bla"
/label=AmpR promoter
CDS 5984..6844
/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(6202..6221)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7015..7603
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7504..7523
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 7703..21
/label=lac promoter
/note="promoter for the E. coli lac operon"
ORIGIN
1 atgcttccgg ctcgtatgtt gaaagaggag aaaggatcta tgaacagtca gcaaaatcct
61 gatttagctg ttcatccctt ggcaattcct atggaaggct tactaggaga aagtgctaca
121 actcttgaga agaatgtaat tgccacacaa ctctcagagg aagcccaagt aaagctagag
181 gtaatccaaa gtttactgga accctgcgat cgcacaactt atgggcaaaa gttgcgggaa
241 gcagcagaga aactaaatgt atcgttgcga acggtacaaa ggttggtgaa aaactgggaa
301 caagatggct tagtcggact cactcaaaca agtagggctg ataaaggaaa acaccgcatt
361 ggtgagtttt gggaaaactt cattaccaaa acctacaagg agggtaacaa gggaagtaaa
421 cgtatgaccc ctaaacaagt tgctctcaga gtcgaggcta aagcccgtga attaaaagac
481 tctaagccgc ccaattacaa aaccgtgtta cgggtattag cacccatttt ggaaaagcaa
541 caaaaagcca agagtatccg cagtcctggt tggagaggaa ctacgctttc ggttaaaacc
601 cgtgaaggaa aagatttatc ggttgattac agtaaccatg tttggcaatg tgaccatacc
661 cgcgtggatg tgttgctggt agatcaacat ggtgaaattt taagtcgtcc ctggctaaca
721 acagtaattg atacttactc tcgttgcatt atgggtatca acttgggctt tgatgcaccc
781 agttctgggg tagtagcatt agcgttacgc catgcaattc taccaaagcg ttacggttcc
841 gagtacaaac tgcattgtga gtggggaacc tatggaaaac cagaacattt ttatactgat
901 ggcggtaaag actttcgctc taaccacttg agtcagattg gggcgcaatt gggatttgtc
961 tgtcatttac gcgatcgccc ttctgaaggt ggagtagtag aacgtccctt caaaacatta
1021 aatgaccaac tattttcaac gcttcctggg tacaccggat ctaatgtgca ggaacgccca
1081 gaagatgcag agaaggacgc aagacttact ttgcgagaac tagaacagtt acttgtgcgt
1141 tacatcgtag atcgttacaa ccaaagtatt gatgcgcgga tgggcgacca aacgcgcttt
1201 gagcgttggg aagcaggatt gcctacagtg ccagtaccaa taccagaacg agatttggat
1261 atttgtttaa tgaagcagtc acggcgcact gtgcaaagag gtggttgttt gcagtttcag
1321 aatttaatgt atcgggggga atatttggca ggttatgccg gagaaactgt caacttaagg
1381 tttgacccca gagacattac aacaattttg gtttatcgcc aggaaaacaa tcaggaagta
1441 tttctgactc gcgctcacgc tcaaggtttg gagacagagc aactggcatt agatgaggct
1501 gaggcagcaa gtcgcagact ccgtaccgca gggaaaacta tcagtaacca atcattattg
1561 caagaagttg ttgaccgcga tgctcttgtc gctaccaaga aaagccgtaa ggagcgtcaa
1621 aaattggaac agactgtttt gcgatctgct gctgttgatg aaagtaatag agaatccttg
1681 ccttctcaaa tagttgaacc agatgaagtg gaatctacag aaacggttca ctctcaatac
1741 gaagacattg aggtgtggga ctatgaacaa cttcgtgaag aatatgggtt ttaaacaatg
1801 acagaagctc aggcgatcgc caagcagttg ggtggggtaa aaccggatga tgagtggtta
1861 caagctgaaa ttgctcgtct caagggtaag agcattgtgc ctttacagca ggtaaaaact
1921 ctccatgatt ggttagatgg caagcgcaag gcaagaaaat cttgccgagt agttggggaa
1981 tcgagaactg gcaagacagt tgcttgtgat gcctacagat acaggcacaa acctcagcag
2041 gaagctggac gacctccaac tgtgcctgtc gtttatattc gacctcacca aaaatgtggc
2101 cccaaggatt tgtttaaaaa gattactgag tacctcaagt atcgggtaac aaaagggact
2161 gtatctgatt ttcgagatag gacgatagaa gtactcaagg gttgtggcgt agagatgcta
2221 attattgatg aagctgaccg tctcaagcct gaaacttttg ctgatgtgcg agatattgcc
2281 gaagatttag gaattgctgt ggtactggta ggaacagacc gtttggatgc ggtaattaag
2341 cgggatgagc aggttctcga acgctttcgg gcgcatcttc gctttggtaa attgtcggga
2401 gaggatttta agaacaccgt agaaatgtgg gaacaaatgg ttttgaaact gccagtatct
2461 tctaatctaa agagcaagga gatgctacgg attctcacgt cagcaactga aggctacatt
2521 ggtcgccttg atgagattct tagggaagct gcaattcgtt ccttatcaag aggattgaag
2581 aagattgaca aggctgtttt acaggaagta gctaaggagt acaaatgata gaagcaccag
2641 atgttaaacc ttggctattc ttgattaaac cctatgaagg ggaaagcctg agccactttc
2701 ttggcaggtt cagacgtgcc aaccatttat ccgcaagtgg attgggtact ttggcaggaa
2761 ttggtgctat agtggcacgt tgggaaagat ttcattttaa tcctcgccct agtcagcaag
2821 aattggaagc gatcgcatct gtagtagaag tggatgctca aaggttagcc cagatgttac
2881 cgcctgctgg agtgggaatg cagcatgagc caattcgctt gtgtggggct tgttatgccg
2941 agtcgccttg tcaccgaatt gaatggcagt acaagtcggt gtggaagtgc gatcgccatc
3001 aactcaagat tttagcaaag tgtccaaact gtcaagcacc ttttaaaatg cctgcgctgt
3061 gggaggatgg gtgctgtcac agatgtagga tgccgtttgc agaaatggca aagctacaga
3121 aggtttgatg ataaaaccag aaaaaggtgt gaaattaact aagtccctga attgatctgg
3181 ttgtccaaaa aatttgtgcg atcgcatggc aagattattc ctactattga tgtggcgtga
3241 gtcggataac ttgctctaat gggatcccgg ttaataccaa tagatcagat atgagtaata
3301 aacattacga cctttacctg tggtattcac tgttggcacg acaacacctt tatcgcgcca
3361 atactgtagc tgacgacgcg aacaattagt gatttgagcc gcttctgtgc ttgtgtagaa
3421 ttttccttcc atagaaatca tcctacagaa gtagtttttt aaatttacat tttttaaaag
3481 cacattatgt attcatgtga tattattttt ttatgagtca aataactatt caagctcgac
3541 ttatttcctt tgaatcaaac cgccaacaac tctggaagtt gatggcagat ttaaacacgc
3601 cgttaattaa cgaactgctt tgccagttag gtcaacaccc cgacttcgag aagtggcaac
3661 aaaagggtaa actcccgtct accgttgtga gccagttatg tcaacctctc aaaactgacc
3721 ctcgctttgc aggtcagccc agccgtttat atatgtcggc aattcatatt gtggactaca
3781 tctacaagtc ctggctggct atacagaaac ggcttcaaca gcagctagat ggaaagacgc
3841 gctggctaga aatgctcaat agcgatgctg aattagtaga acttagtggt gacactttag
3901 aggctattcg tgtcaaagct gctgaaattt tggcaatagc tatgccagca tctgagtcag
3961 atagcgcttc acctaaaggg aaaaaaggta aaaaggagaa aaaaccctca tcttctagcc
4021 ctaagcgtag tttatccaag acattatttg acgcttacca agaaacggaa gatatcaaga
4081 gccgtagcgc catcagctac ctgttaaaaa atggctgcaa acttactgac aaagaagaag
4141 attcagaaaa atttgctaaa cgtcgtcgtc aagttgaaat ccaaattcaa aggcttaccg
4201 aaaagttaat aagtcggatg cctaaaggtc gagatttgac caatgctaaa tggttggaga
4261 cactcttgac tgctacaacc actgttgctg aagacaacgc ccaagccaaa cgctggcagg
4321 atattctgtt aactcgatca agttctctcc cattccccct tgtttttgaa accaacgagg
4381 atatggtttg gtcaaagaat caaaagggta ggctgtgtgt tcacttcaat ggcttaagcg
4441 atttaatttt tgaggtgtac tgcggcaatc gtcaacttca ctggtttcaa cgcttcctag
4501 aagaccaaca gactaaacgc aaaagcaaaa atcagcattc tagcggcttg ttcacactca
4561 gaaatggtca tctagtttgg cttgaaggtg agggtaaagg ggaaccttgg aatcttcacc
4621 acttgaccct ttactgctgt gttgacaatc gcttgtggac agaggaggga acagaaatcg
4681 ttcgccaaga gaaagcagat gaaattacta aattcatcac aaacatgaag aagaaaagcg
4741 atctaagcga tacacagcaa gctttgattc aacgtaaaca atcaacactt actcgaataa
4801 acaattcctt tgagcgtcct agccaacccc tttatcaagg tcaatcacac attttggttg
4861 gagtaagcct gggactagaa aaacctgcca cagtagcagt agtagatgcg atcgccaaca
4921 aagtcttggc ttaccggagt attaaacaat tacttggcga caattacgaa ctgctaaatc
4981 gccagagacg acaacagcag tacctatctc acgaacgcca caaagcacaa aaaaacttct
5041 ctcccaatca atttggagca tctgagttag ggcaacatat agacagatta ttagctaaag
5101 caattgtagc gttagcgaga acctacaaag ctggcagtat tgtcttgccc aagttagggg
5161 atatgcggga ggttgtccaa agtgaaattc aagctatagc agaacaaaaa tttcccggtt
5221 atattgaagg tcagcaaaaa tatgccaaac agtaccgggt taatgttcat cggtggagct
5281 acggcagatt aattcaaagc attcaaagta aagcagctca aacaggaatt gtgattgagg
5341 agggaaaaca acctattcga ggtagtcccc acgacaaagc aaaggaatta gcactttctg
5401 cttacaatct ccgcctaact aggcgaagtt aagtaaggat ctccaggcat caaataaaac
5461 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
5521 tctactagag agaccctgac agctagctca gtcctaggta taatgctagc atattaatag
5581 cgccgcaatt catgctgctt gcagcctctg aattttgtta aatgagggtt agtttgactg
5641 tataaataca gtcttgcttt ctgaccctgg tagctgctca ccctgatgct gctgtcaata
5701 gacaggatag gtgcgctccc agcaataagg gcgcggatgt actgctgtag tggctactga
5761 atcacccccg atcaaggggg aaccctaaat gggttgaaag gaagagcgag atctcgagct
5821 cttcggctca ccttcgggtg ggcctttctg cgcaggtggc acttttcggg gaaatgtgcg
5881 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
5941 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
6001 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
6061 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
6121 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
6181 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
6241 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
6301 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
6361 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
6421 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
6481 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
6541 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
6601 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
6661 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
6721 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
6781 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
6841 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
6901 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
6961 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
7021 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
7081 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
7141 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
7201 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
7261 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
7321 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
7381 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
7441 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
7501 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
7561 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
7621 ctttttacgg ttcctggcct tttgctggcc ttttgctcaa cgccaagctc caaaacgatc
7681 tcaagaagat catcttatta attttacact tt
//
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