| 质粒名称: | pAW212 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 113234 |
| 存储实验室: | Jennifer Doudna |
|
载体骨架基本信息 |
|
| 载体名称: | pCDF1b |
| 载体出品公司: | -- |
| 空载体大小: | 4000 bp |
| 完整质粒大小: | 4909 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: |
链霉素50 μg/mL
|
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Cas1 |
| 物种来源: | E. coli |
| 插入大小: | 924 bp |
| 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: | pAW212 was a gift from Jennifer Doudna (Addgene plasmid # 113234 ; http://n2t.net/addgene:113234 ; RRID:Addgene_113234) |
| 参考文献部分: | Structures of the CRISPR genome integration complex. Wright AV, Liu JJ, Knott GJ, Doxzen KW, Nogales E, Doudna JA. Science. 2017 Sep 15;357(6356):1113-1118. doi: 10.1126/science.aao0679. Epub 2017 Jul 20. 10.1126/science.aao0679 PubMed 28729350 |
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 pAW212 4909 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION E. coli expression vector for untagged E. coli Cas1 + Cas2 with
CRISPR array, leader GGTAG->CCATC.
ACCESSION .
VERSION .
KEYWORDS pAW212
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4909)
AUTHORS Wright AV, Liu JJ, Knott GJ, Doxzen KW, Nogales E, Doudna JA
TITLE Structures of the CRISPR genome integration complex.
JOURNAL Science. 2017 Sep 15;357(6356):1113-1118. doi:
10.1126/science.aao0679. Epub 2017 Jul 20.
PUBMED 28729350
REFERENCE 2 (bases 1 to 4909)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4909
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 3..27
/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)."
CDS 1313..1357
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
primer_bind complement(1395..1413)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1409..1456
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(1627..2418)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
promoter complement(2419..2510)
/gene="bla"
/label=AmpR promoter
rep_origin complement(2558..3296)
/direction=LEFT
/label=CloDF13 ori
/note="Plasmids containing the CloDF13 (CDF) origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
CDS complement(3687..4769)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 4731..4750
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4770..4847)
/gene="lacI"
/label=lacI promoter
primer_bind 4893..3
/label=T7
/note="T7 promoter, forward primer"
promoter 4893..2
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
61 gagatatacc atggcgatga cctggcttcc ccttaatccc attccactca aagatcgcgt
121 ctccatgatc tttctgcaat atgggcagat cgatgtaata gatggcgcgt ttgtacttat
181 cgacaagaca gggatccgca ctcatattcc tgttggctcg gttgcctgca tcatgctgga
241 acctggtaca cgggtttcgc atgcagctgt acgcctggct gcgcaagttg gaacattgtt
301 ggtatgggtg ggggaagcgg gcgttcgtgt ttatgcttct ggtcagcctg gaggtgcgcg
361 ttcagataag ctgctctatc aggcaaaact tgctctggat gaagatttgc gtctgaaggt
421 cgtacgtaaa atgtttgaac ttcggtttgg agaacctgcg cctgcccggc gctccgtaga
481 gcaactcaga ggtatagaag gcagtcgcgt gcgggcaacc tacgcacttc tggcgaagca
541 atacggcgtg acatggaatg gacgtcgcta cgatccgaaa gactgggaaa agggcgatac
601 gatcaaccaa tgcattagcg ctgcaacttc ctgtttatac ggcgtaactg aagcggcgat
661 acttgcagct ggttatgcac cagctattgg gtttgtgcat acaggaaagc ctctttcctt
721 tgtttacgat attgcagaca tcattaaatt tgacactgtt gtaccgaaag cttttgagat
781 agcgcgtcgt aaccctggtg agccggaccg ggaagtccgt ttggcgtgca gggatatttt
841 tcgcagtagt aaaacattag ccaaattgat tccgcttata gaggacgtgc ttgccgctgg
901 agaaatacaa ccgccggccc cacctgaaga tgcacagcct gttgccattc cgcttcctgt
961 ttcactggga gatgcaggcc atcgctcgag ctgaaggaga tataccatga gtatgttggt
1021 cgtggtcact gaaaatgtac ctccgcgctt acgaggcaga ttagccatct ggttgttgga
1081 ggtacgtgca ggggtatatg taggtgatgt atccgcaaaa attcgtgaaa tgatctggga
1141 acaaatagct ggactggcgg aagaaggcaa tgtagtgatg gcatgggcaa cgaatacgga
1201 aacgggattt gagttccaga catttgggtt aaacaggcgt accccggtag atttggatgg
1261 tttaaggttg gtgtcttttt tacctgtttg agcggccgca ctcgagtctg gtaaagaaac
1321 cgctgctgcg aaatttgaac gccagcacat ggactcgtct actagcgcag cttaattaac
1381 ctaggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
1441 cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca cactgcttcc
1501 ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc
1561 tgaaccgacg accgggtcat cgtggccgga tcttgcggcc cctcggcttg aacgaattgt
1621 tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga caaattcttc
1681 caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag cctgtctagc
1741 ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt cggcagcgac
1801 atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca acgtaagcac
1861 tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta aggtttcatt
1921 tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg ccgctggacc
1981 taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat caatgtcgat
2041 cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc caaattgcag
2101 ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc
2161 tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat
2221 caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca aatcaatatc
2281 actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg ccagcaacgt
2341 cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg atacttcggc
2401 gatcaccgct tccctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
2461 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa tagctagctc
2521 actcggtcgc tacgctccgg gcgtgagact gcggcgggcg ctgcggacac atacaaagtt
2581 acccacagat tccgtggata agcaggggac taacatgtga ggcaaaacag cagggccgcg
2641 ccggtggcgt ttttccatag gctccgccct cctgccagag ttcacataaa cagacgcttt
2701 tccggtgcat ctgtgggagc cgtgaggctc aaccatgaat ctgacagtac gggcgaaacc
2761 cgacaggact taaagatccc caccgtttcc ggcgggtcgc tccctcttgc gctctcctgt
2821 tccgaccctg ccgtttaccg gatacctgtt ccgcctttct cccttacggg aagtgtggcg
2881 ctttctcata gctcacacac tggtatctcg gctcggtgta ggtcgttcgc tccaagctgg
2941 gctgtaagca agaactcccc gttcagcccg actgctgcgc cttatccggt aactgttcac
3001 ttgagtccaa cccggaaaag cacggtaaaa cgccactggc agcagccatt ggtaactggg
3061 agttcgcaga ggatttgttt agctaaacac gcggttgctc ttgaagtgtg cgccaaagtc
3121 cggctacact ggaaggacag atttggttgc tgtgctctgc gaaagccagt taccacggtt
3181 aagcagttcc ccaactgact taaccttcga tcaaaccacc tccccaggtg gttttttcgt
3241 ttacagggca aaagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
3301 ctactgaacc gctctagata cctttttaat caatggatta agtactcttt aacataatgg
3361 atgtgttgtt tgtgtgatac tataaagttc catcattgtg actggcttaa aaaatcatta
3421 attaataata ggttatgttt agagtgttcc ccgcgccagc ggggataaac cgagcacaaa
3481 tatcatcgct caatctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
3541 agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg
3601 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
3661 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
3721 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
3781 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
3841 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
3901 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
3961 accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
4021 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
4081 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
4141 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
4201 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
4261 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
4321 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
4381 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
4441 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
4501 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
4561 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
4621 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
4681 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
4741 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
4801 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
4861 ctctccctta tgcgactcct gcattaggaa attaatacga ctcactata
//
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