| 质粒名称: | pFS_0453_pET30_pCat-tetR-Term-ptetA-FsRT-Cas1(opt)-Cas2(opt)_T7term_ARRAY2_FaqI |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 117006 |
| 存储实验室: | Randall Platt |
|
载体骨架基本信息 |
|
| 载体名称: | pET30b+ |
| 载体出品公司: | Novagen |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Fusicatenibacter saccharivorans RT-Cas1-Cas2 |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | T7 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | T7 |
| 3' 测序引物 | T7-term |
|
文献引用方法 |
|
| 材料和方法部分: | pFS_0453_pET30_pCat-tetR-Term-ptetA-FsRT-Cas1(opt)-Cas2(opt)_T7term_ARRAY2_FaqI was a gift from Randall Platt (Addgene plasmid # 117006 ; http://n2t.net/addgene:117006 ; RRID:Addgene_117006) |
| 参考文献部分: | Transcriptional recording by CRISPR spacer acquisition from RNA. Schmidt F, Cherepkova MY, Platt RJ. Nature. 2018 Oct;562(7727):380-385. doi: 10.1038/s41586-018-0569-1. Epub 2018 Oct 3. 10.1038/s41586-018-0569-1 PubMed 30283135 |
pFS_0453_pET30_pCat-tetR-Term-ptetA-FsRT-Cas1(opt)-Cas2(opt)
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
-Cas2(opt)_T7term_ARRAY2_FaqI质粒图谱.png)
载体简介
载体序列
LOCUS pFS_0453_pET30_pCat-tetR-Term-ptetA-FsRT-Cas1(opt)-Cas2(opt)_T7term_ARRAY2_FaqI 6346 bp ds-DNA circular SYN 02-NOV-2018
DEFINITION Expresses E. coli codon-optimized FsRT-Cas1-Cas2 under ptetA
promoter, encodes FsCRISPR array 2, compatible with SENECA
acquisition readout.
ACCESSION .
VERSION .
KEYWORDS
pFS_0453_pET30_pCat-tetR-Term-ptetA-FsRT-Cas1(opt)-Cas2(o...
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6346)
TITLE Platt lab unpublished CRISPR plasmids
REFERENCE 2 (bases 1 to 6346)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 2, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6346
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(210..1025)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 933..952
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 1017..1036
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 1147..1735
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1636..1655
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1889..1906
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 1921..2060
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(1986..2005)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2146..2168
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2162..2353)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
RBS 2883..2894
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 2901..3524
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind complement(2974..2993)
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
terminator 3569..3640
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3656..3683
/label=T7Te terminator
/note="phage T7 early transcription terminator"
protein_bind 3698..3716
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3723..3741
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind complement(6025..6043)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 6039..6086
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 ggccgctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
61 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
121 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
181 caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac
241 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
301 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
361 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
421 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
481 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
541 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
601 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
661 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
721 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
781 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
841 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
901 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
961 tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg
1021 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa
1081 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
1141 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
1201 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
1261 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
1321 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
1381 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
1441 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
1501 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
1561 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
1621 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
1681 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
1741 cagcaacgcg gcctttttac ggttcctggt cttttgctgg ccttttgctc acatgttctt
1801 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
1861 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
1921 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
1981 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
2041 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
2101 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
2161 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
2221 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
2281 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
2341 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
2401 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
2461 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
2521 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
2581 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
2641 gactttacga aacacggaaa ccgaagatca ttcatgttgt tgctcaggtc gcagacgttt
2701 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
2761 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
2821 gggccgccat gccggcggca cgtaagaggt tccaactttc accataatga aacatactag
2881 agaaagagga gaaatactag atgtccagat tagataaaag taaagtgatt aacagcgcat
2941 tagagctgct taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc
3001 taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg
3061 ccttagccat tgagatgtta gataggcacc atactcactt ttgcccttta gaaggggaaa
3121 gctggcaaga ttttttacgt aataacgcta aaagttttag atgtgcttta ctaagtcatc
3181 gcgatggagc aaaagtacat ttaggtacac ggcctacaga aaaacagtat gaaactctcg
3241 aaaatcaatt agccttttta tgccaacaag gtttttcact agagaatgca ttatatgcac
3301 tcagcgctgt ggggcatttt actttaggtt gcgtattgga agatcaagag catcaagtcg
3361 ctaaagaaga aagggaaaca cctactactg atagtatgcc gccattatta cgacaagcta
3421 tcgaattatt tgatcaccaa ggtgcagagc cagccttctt attcggcctt gaattgatca
3481 tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc ctaataacac tgatagtgct
3541 agtgtagatc actactagag ccaggcatca aataaaacga aaggctcagt cgaaagactg
3601 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc
3661 accttcgggt gggcctttct gcgtttatat actagagtcc ctatcagtga tagagattga
3721 catccctatc agtgatagag atactgagca ctactagaga agcttgcgga ggagcatgca
3781 tgtttaccat cgacgagatg ctgagcaaga acaaccaacg tctggcgttc gaacactttg
3841 cgaccaaaaa cgacggttgc ggcccggacg gtatgcacgt tagcgaactg gagaagtact
3901 ggcgtatgaa tcacgaccag atcattagcg atctgaagaa ccaggaatac cagccgggta
3961 ttatcctgat tcgtgagcac atgaataaaa ccggcaaacg ccgtaatatc gcgagcctga
4021 acgttattga tcgtttcatt acccgtctgc tgagccagaa actgaaccgt tatctggcgc
4081 cgattttctg cgagaacagc tatgcgtacc aggacagcaa aggcgttatg ccggcggttc
4141 tgaaagcgaa ggagtacgtt gagctgggta tgcgtcacgt gattgaaatc gatctgaaaa
4201 actactttga caccattccg ctggaaaacc tgatcccgga gatcgaacgc tacatcaccg
4261 atgaggcggt tctgcatctg atcaagcaat acctgttttg cgatattagc ttcgagggca
4321 aaatcagccg taaaacccaa ggtatcgttc aaggtaacgc gatcagcccg atcctgagca
4381 acctgtacct gaacgacttc gacaaggaac tggacgagag caagctgtgc tggattcgtt
4441 acgcggacaa tatctacatc tatatggata gctatgagaa ggcgctgctg gtgtatagcg
4501 agctgaccga gcgtctggag cgtcgcaagc tgaccgttaa caaggagaaa agcggcgttt
4561 ttgatgtgag cacccgtagc attctgggct acgacatcct gattcgtaac aagaaagtgg
4621 acgttcgtaa gcatatttac aaaagcgtta atcagtacag caactggcac gatagccgcc
4681 tggagtttat caacggtcgt taccacatca ccagcgatgg catcctgaat cgccaagact
4741 tcggcctgct gttcgagaat gagcagaaaa aacactacat cccggttgag gttagcgatc
4801 agctgaacat ctacggtaat gttaccctgg cgagcaatgt gctgcaaagt ttcagcaacc
4861 gcgaaatcaa agtgagcttc ttcgacaagt atggtcgtct gattggtagc ttcctgccgg
4921 aaaaaaccaa gaaaagcgcg gagatcattc tggtgcagag caagaactat ctgaacgaag
4981 atgtgcgtat ggataccgcg cgccgtatgg aaattgcggg tctgcacaac atccgtgcga
5041 atctgcgcta ctacgataag aaacacaagg gcgactttaa agagaaggtt gacgcgatta
5101 gcggctacat tgacgcgctg aaccgtgcgc cgagcgttaa tgatatgatg ctgctggaag
5161 cgaaagcgcg ccaactgtat tatacctgtt ttaaccaaat cctggaaacc agcgacttcc
5221 agttcgaaaa gcgtaccaag cgtccgccga aagatgcgat caatgcgtgc attagcttcg
5281 gcaataccct gctgtataat ctatttgtga atattatctg gaagaagggt ctggacccgc
5341 gctttggcgt tgttcacgcg agcaacaaac gcaatcaaag cctgaacctg gacttcgcgg
5401 acatctttaa accgatcgtg attgaccgca ttatcttcac catgatcaat aagaaaatgc
5461 tgaccctgct gaccgatttt gaaaccagca accagggtgt gtatctgagc cgtgagggca
5521 agaacatctt cctgcaaatg tacgaggaga agctgaaaag ccgcattacc atcaagggta
5581 aagagatgag ctattaccag ctgctggaga gcgaagtgca gaattacaag aatttcattc
5641 tgaccggcga aacctataaa ccgtataagt attattaacg tacgttaact ttaagaagga
5701 gaacttaaga tgtatgttat cctggtgtac gacatccacc agaagcgtgt tggtaaagcg
5761 ctgaaaatct gccgtaagta cctgatccac atccaaaaga gcgtgttcga aggcaacatc
5821 accgagagca aactgaaggc gctgaaagag gaactgggtc acctgattga cacccagatg
5881 gatagcgtga ttatctacca cctggacagc gttaagtaca ccaaaaagga gcagatcggt
5941 attgtgcaga gcaccagcaa tgtgatctaa ctcgagatcc ggctgctaac aaagcccgaa
6001 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
6061 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga taccaggtgc
6121 ggggaagctt atgttccata gcaaaaagtc ggtcagtctc gtggctgaaa tcatgagttc
6181 cacaaaatgg ctgaaattca aggaaaatca ggaatctcag aaaaacgatc gaccgacttt
6241 ttcgataaaa tggttgcaaa aatgagaaaa atctgattta atagaatctg aaaacagcgg
6301 aaatgctgtt gtcgtacttt acctaaaagg aattgaaacg tcccgc
//
