| 质粒名称: | pHelper_ShCAST |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 127922 |
| 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 |
| 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 was a gift from Feng Zhang (Addgene plasmid # 127922 ; http://n2t.net/addgene:127922 ; RRID:Addgene_127922) |
| 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
pHelperShCAST载体基本信息
pHelperShCAST载体图谱质粒图谱和多克隆位点信息
pHelperShCAST载体简介
pHelperShCAST载体序列
LOCUS pHelper_ShCAST 8150 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses ShCAST and a crRNA targeting PSP1.
ACCESSION .
VERSION .
KEYWORDS pHelper_ShCAST
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8150)
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 8150)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8150
/organism="synthetic DNA construct"
/mol_type="other DNA"
RBS 1..12
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
promoter 5857..5891
/label=J23119 promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119)"
primer_bind complement(6031..6050)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6150..6172
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6210..6228)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6296..6400
/gene="bla"
/label=AmpR promoter
CDS 6401..7261
/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(6619..6638)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7432..8020
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7921..7940
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 8120..8150
/label=lac promoter
/note="promoter for the E. coli lac operon"
ORIGIN
1 aaagaggaga aaggatctat gaacagtcag caaaatcctg atttagctgt tcatcccttg
61 gcaattccta tggaaggctt actaggagaa agtgctacaa ctcttgagaa gaatgtaatt
121 gccacacaac tctcagagga agcccaagta aagctagagg taatccaaag tttactggaa
181 ccctgcgatc gcacaactta tgggcaaaag ttgcgggaag cagcagagaa actaaatgta
241 tcgttgcgaa cggtacaaag gttggtgaaa aactgggaac aagatggctt agtcggactc
301 actcaaacaa gtagggctga taaaggaaaa caccgcattg gtgagttttg ggaaaacttc
361 attaccaaaa cctacaagga gggtaacaag ggaagtaaac gtatgacccc taaacaagtt
421 gctctcagag tcgaggctaa agcccgtgaa ttaaaagact ctaagccgcc caattacaaa
481 accgtgttac gggtattagc acccattttg gaaaagcaac aaaaagccaa gagtatccgc
541 agtcctggtt ggagaggaac tacgctttcg gttaaaaccc gtgaaggaaa agatttatcg
601 gttgattaca gtaaccatgt ttggcaatgt gaccataccc gcgtggatgt gttgctggta
661 gatcaacatg gtgaaatttt aagtcgtccc tggctaacaa cagtaattga tacttactct
721 cgttgcatta tgggtatcaa cttgggcttt gatgcaccca gttctggggt agtagcatta
781 gcgttacgcc atgcaattct accaaagcgt tacggttccg agtacaaact gcattgtgag
841 tggggaacct atggaaaacc agaacatttt tatactgatg gcggtaaaga ctttcgctct
901 aaccacttga gtcagattgg ggcgcaattg ggatttgtct gtcatttacg cgatcgccct
961 tctgaaggtg gagtagtaga acgtcccttc aaaacattaa atgaccaact attttcaacg
1021 cttcctgggt acaccggatc taatgtgcag gaacgcccag aagatgcaga gaaggacgca
1081 agacttactt tgcgagaact agaacagtta cttgtgcgtt acatcgtaga tcgttacaac
1141 caaagtattg atgcgcggat gggcgaccaa acgcgctttg agcgttggga agcaggattg
1201 cctacagtgc cagtaccaat accagaacga gatttggata tttgtttaat gaagcagtca
1261 cggcgcactg tgcaaagagg tggttgtttg cagtttcaga atttaatgta tcggggggaa
1321 tatttggcag gttatgccgg agaaactgtc aacttaaggt ttgaccccag agacattaca
1381 acaattttgg tttatcgcca ggaaaacaat caggaagtat ttctgactcg cgctcacgct
1441 caaggtttgg agacagagca actggcatta gatgaggctg aggcagcaag tcgcagactc
1501 cgtaccgcag ggaaaactat cagtaaccaa tcattattgc aagaagttgt tgaccgcgat
1561 gctcttgtcg ctaccaagaa aagccgtaag gagcgtcaaa aattggaaca gactgttttg
1621 cgatctgctg ctgttgatga aagtaataga gaatccttgc cttctcaaat agttgaacca
1681 gatgaagtgg aatctacaga aacggttcac tctcaatacg aagacattga ggtgtgggac
1741 tatgaacaac ttcgtgaaga atatgggttt taaacaatga cagaagctca ggcgatcgcc
1801 aagcagttgg gtggggtaaa accggatgat gagtggttac aagctgaaat tgctcgtctc
1861 aagggtaaga gcattgtgcc tttacagcag gtaaaaactc tccatgattg gttagatggc
1921 aagcgcaagg caagaaaatc ttgccgagta gttggggaat cgagaactgg caagacagtt
1981 gcttgtgatg cctacagata caggcacaaa cctcagcagg aagctggacg acctccaact
2041 gtgcctgtcg tttatattcg acctcaccaa aaatgtggcc ccaaggattt gtttaaaaag
2101 attactgagt acctcaagta tcgggtaaca aaagggactg tatctgattt tcgagatagg
2161 acgatagaag tactcaaggg ttgtggcgta gagatgctaa ttattgatga agctgaccgt
2221 ctcaagcctg aaacttttgc tgatgtgcga gatattgccg aagatttagg aattgctgtg
2281 gtactggtag gaacagaccg tttggatgcg gtaattaagc gggatgagca ggttctcgaa
2341 cgctttcggg cgcatcttcg ctttggtaaa ttgtcgggag aggattttaa gaacaccgta
2401 gaaatgtggg aacaaatggt tttgaaactg ccagtatctt ctaatctaaa gagcaaggag
2461 atgctacgga ttctcacgtc agcaactgaa ggctacattg gtcgccttga tgagattctt
2521 agggaagctg caattcgttc cttatcaaga ggattgaaga agattgacaa ggctgtttta
2581 caggaagtag ctaaggagta caaatgatag aagcaccaga tgttaaacct tggctattct
2641 tgattaaacc ctatgaaggg gaaagcctga gccactttct tggcaggttc agacgtgcca
2701 accatttatc cgcaagtgga ttgggtactt tggcaggaat tggtgctata gtggcacgtt
2761 gggaaagatt tcattttaat cctcgcccta gtcagcaaga attggaagcg atcgcatctg
2821 tagtagaagt ggatgctcaa aggttagccc agatgttacc gcctgctgga gtgggaatgc
2881 agcatgagcc aattcgcttg tgtggggctt gttatgccga gtcgccttgt caccgaattg
2941 aatggcagta caagtcggtg tggaagtgcg atcgccatca actcaagatt ttagcaaagt
3001 gtccaaactg tcaagcacct tttaaaatgc ctgcgctgtg ggaggatggg tgctgtcaca
3061 gatgtaggat gccgtttgca gaaatggcaa agctacagaa ggtttgatga taaaaccaga
3121 aaaaggtgtg aaattaacta agtccctgaa ttgatctggt tgtccaaaaa atttgtgcga
3181 tcgcatggca agattattcc tactattgat gtggcgtgag tcggataact tgctctaatg
3241 ggatcccggt taataccaat agatcagata tgagtaataa acattacgac ctttacctgt
3301 ggtattcact gttggcacga caacaccttt atcgcgccaa tactgtagct gacgacgcga
3361 acaattagtg atttgagccg cttctgtgct tgtgtagaat tttccttcca tagaaatcat
3421 cctacagaag tagtttttta aatttacatt ttttaaaagc acattatgta ttcatgtgat
3481 attatttttt tatgagtcaa ataactattc aagctcgact tatttccttt gaatcaaacc
3541 gccaacaact ctggaagttg atggcagatt taaacacgcc gttaattaac gaactgcttt
3601 gccagttagg tcaacacccc gacttcgaga agtggcaaca aaagggtaaa ctcccgtcta
3661 ccgttgtgag ccagttatgt caacctctca aaactgaccc tcgctttgca ggtcagccca
3721 gccgtttata tatgtcggca attcatattg tggactacat ctacaagtcc tggctggcta
3781 tacagaaacg gcttcaacag cagctagatg gaaagacgcg ctggctagaa atgctcaata
3841 gcgatgctga attagtagaa cttagtggtg acactttaga ggctattcgt gtcaaagctg
3901 ctgaaatttt ggcaatagct atgccagcat ctgagtcaga tagcgcttca cctaaaggga
3961 aaaaaggtaa aaaggagaaa aaaccctcat cttctagccc taagcgtagt ttatccaaga
4021 cattatttga cgcttaccaa gaaacggaag atatcaagag ccgtagcgcc atcagctacc
4081 tgttaaaaaa tggctgcaaa cttactgaca aagaagaaga ttcagaaaaa tttgctaaac
4141 gtcgtcgtca agttgaaatc caaattcaaa ggcttaccga aaagttaata agtcggatgc
4201 ctaaaggtcg agatttgacc aatgctaaat ggttggagac actcttgact gctacaacca
4261 ctgttgctga agacaacgcc caagccaaac gctggcagga tattctgtta actcgatcaa
4321 gttctctccc attccccctt gtttttgaaa ccaacgagga tatggtttgg tcaaagaatc
4381 aaaagggtag gctgtgtgtt cacttcaatg gcttaagcga tttaattttt gaggtgtact
4441 gcggcaatcg tcaacttcac tggtttcaac gcttcctaga agaccaacag actaaacgca
4501 aaagcaaaaa tcagcattct agcggcttgt tcacactcag aaatggtcat ctagtttggc
4561 ttgaaggtga gggtaaaggg gaaccttgga atcttcacca cttgaccctt tactgctgtg
4621 ttgacaatcg cttgtggaca gaggagggaa cagaaatcgt tcgccaagag aaagcagatg
4681 aaattactaa attcatcaca aacatgaaga agaaaagcga tctaagcgat acacagcaag
4741 ctttgattca acgtaaacaa tcaacactta ctcgaataaa caattccttt gagcgtccta
4801 gccaacccct ttatcaaggt caatcacaca ttttggttgg agtaagcctg ggactagaaa
4861 aacctgccac agtagcagta gtagatgcga tcgccaacaa agtcttggct taccggagta
4921 ttaaacaatt acttggcgac aattacgaac tgctaaatcg ccagagacga caacagcagt
4981 acctatctca cgaacgccac aaagcacaaa aaaacttctc tcccaatcaa tttggagcat
5041 ctgagttagg gcaacatata gacagattat tagctaaagc aattgtagcg ttagcgagaa
5101 cctacaaagc tggcagtatt gtcttgccca agttagggga tatgcgggag gttgtccaaa
5161 gtgaaattca agctatagca gaacaaaaat ttcccggtta tattgaaggt cagcaaaaat
5221 atgccaaaca gtaccgggtt aatgttcatc ggtggagcta cggcagatta attcaaagca
5281 ttcaaagtaa agcagctcaa acaggaattg tgattgagga gggaaaacaa cctattcgag
5341 gtagtcccca cgacaaagca aaggaattag cactttctgc ttacaatctc cgcctaacta
5401 ggcgaagtta acaaatatct gaaccttgat aatagaatat taatagcgcc gcaattcatg
5461 ctgcttgcag cctctgaatt ttgttaaatg agggttagtt tgactgtata aatacagtct
5521 tgctttctga ccctggtagc tgctcaccct gatgctgctg tcaatagaca ggataggtgc
5581 gctcccagca ataagggcgc ggatgtactg ctgtagtggc tactgaatca cccccgatca
5641 agggggaacc ctccccaatt cttcatttga aggactaaaa tcaaggcaaa atttctaaga
5701 gatccgcgca agttccaaat accttatctc gtcttgattt catctttttt taaccaaggc
5761 atgattcttg agactgaggc tcaaatgaga aattgggaaa catccgcgct gaagatatct
5821 ggaaaggttg gctcacaaca ttttgtaact gggtagttga cagctagctc agtcctaggt
5881 ataatgctag cgtggcaaca accttccagg tactaggtgg gttgaaagga gaagtcattt
5941 aataaggcca ctgttaaaag tggcaacaac cttccaggta ctaggtgggt tgaaagcggc
6001 cgacgcgctg ggctactggt gcactctcag tacaatctgc tctgatgccg catagttaag
6061 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
6121 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
6181 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
6241 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
6301 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
6361 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
6421 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
6481 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
6541 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
6601 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
6661 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
6721 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
6781 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
6841 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
6901 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
6961 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
7021 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
7081 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
7141 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
7201 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
7261 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
7321 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
7381 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
7441 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
7501 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
7561 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
7621 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
7681 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
7741 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
7801 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
7861 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
7921 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
7981 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
8041 tttacggttc ctggcctttt gctggccttt tgctcaacgc caagctccaa aacgatctca
8101 agaagatcat cttattaatt ttacacttta tgcttccggc tcgtatgttg
//
