| 质粒名称: | pFS_0235-pET30-Fusicatenibacter_saccharivorans_ARRAY2_FaqI |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 116954 |
| 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_0235-pET30-Fusicatenibacter_saccharivorans_ARRAY2_FaqI was a gift from Randall Platt (Addgene plasmid # 116954 ; http://n2t.net/addgene:116954 ; RRID:Addgene_116954) |
| 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_0235-pET30-Fusicatenibacter_saccharivorans_ARRAY2_FaqI
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 pFS_0235-pET30-Fusicatenibacter_saccharivorans_ARRAY2_FaqI 7289 bp ds-DNA circular SYN 02-NOV-2018
DEFINITION Expresses FsRT-Cas1-Cas2 under pT7lac promoter, encodes FsCRISPR
array 2, compatible with SENECA acquisition readout.
ACCESSION .
VERSION .
KEYWORDS pFS_0235-pET30-Fusicatenibacter_saccharivorans_ARRAY2_FaqI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7289)
TITLE Platt lab unpublished CRISPR plasmids
REFERENCE 2 (bases 1 to 7289)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 2, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7289
/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"
CDS complement(3162..4244)
/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 4206..4225
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4245..4322)
/gene="lacI"
/label=lacI promoter
primer_bind 4528..4547
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 4635..4654
/label=T7
/note="T7 promoter, forward primer"
promoter 4635..4653
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 4654..4678
/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)."
RBS 4693..4715
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
primer_bind complement(6968..6986)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 6982..7029
/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 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
2881 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
2941 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
3001 gtcctacgag ttgcatgata aagaagatag tcataagtgc ggcgacgata gtcatgcccc
3061 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
3121 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
3181 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
3241 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
3301 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
3361 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct atcttcggta
3421 tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg
3481 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
3541 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
3601 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
3661 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
3721 accagatgct ccacgcccag tcgcgtacca tcttcatggg agaaaataat actgttgatg
3781 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
3841 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
3901 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
3961 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
4021 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
4081 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
4141 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
4201 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
4261 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
4321 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
4381 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
4441 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
4501 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
4561 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatcgatct
4621 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
4681 tctagaaata attttgttta actttaagaa ggagatatac atatgttcac tatagacgag
4741 atgctatcaa agaacaacca aagattggct ttcgagcact tcgctactaa gaacgacgga
4801 tgtggccctg atggtatgca cgtatctgag ctggagaaat attggcgtat gaatcacgat
4861 cagatcatct ccgacttgaa gaaccaagag taccaacccg gcattatttt gattagagaa
4921 cacatgaaca aaacaggtaa gagaaggaac attgcatctc tcaacgtgat cgacaggttc
4981 ataactcgac tgctgtcgca aaagttgaat agatacttag cccccatttt ctgcgagaac
5041 tcatacgctt accaagactc aaaaggggtg atgcccgccg tactaaaagc caaggagtac
5101 gtcgagttgg gtatgagaca cgttatcgag atagacctca agaattactt cgacacgatc
5161 ccgttagaga acttaatccc agagatcgag cgttacataa cagatgaggc agtgctgcac
5221 ttgattaaac agtacttatt ctgtgacatc tcgttcgagg gtaagatatc acgtaaaact
5281 caagggatcg ttcagggtaa cgcaatttcc ccaatactat ccaacctcta cctgaacgac
5341 ttcgacaagg agctggacga gtcgaagctg tgctggatca gatacgccga caacatatac
5401 atctatatgg acagctacga gaaggcccta ctggtttatt ccgagcttac cgagagactt
5461 gagagaagga aattaaccgt taacaaggag aaatcagggg tgtttgatgt tagtacccgt
5521 tctatactag gttacgacat cctgatcaga aacaagaagg ttgacgtgcg taaacacatc
5581 tacaagtctg ttaaccagta ctccaactgg cacgactctc gattggagtt catcaacggt
5641 cgttaccaca tcacttccga cggtatcctc aatagacaag acttcggtct actgttcgag
5701 aatgagcaga agaagcacta catacctgtt gaggtttctg accagttgaa tatttacgga
5761 aacgttacgc tcgccagcaa cgttcttcaa tcattctcca accgagagat caaagtgtcc
5821 ttcttcgaca agtacgggag acttattgga tctttcctgc ccgagaaaac taagaaaagc
5881 gccgagataa tactggttca gtctaagaac taccttaacg aggacgttag gatggacacg
5941 gcccgacgta tggagatcgc cggtttgcat aacatcagag ccaacctcag atactacgac
6001 aagaagcaca aaggtgactt taaggaaaag gtagacgcca tttctggcta tattgacgcc
6061 ctcaatagag caccatccgt gaacgacatg atgctcttag aggctaaagc tcgtcagctg
6121 tactacacat gcttcaacca gatcttagag acatccgact ttcagttcga gaagcgtacc
6181 aagcgaccac caaaagacgc catcaacgcc tgtatttcat tcggcaacac attactgtac
6241 aacttgtttg tgaatatcat ttggaagaag ggacttgatc caaggttcgg ggttgttcac
6301 gccagcaaca agcgtaatca gtcccttaac ctggattttg ctgacatctt caagcctata
6361 gtcatagata ggatcatctt cacaatgatc aacaagaaga tgctgacctt gttgacggac
6421 ttcgagactt ctaaccaggg agtttacctt tcccgagagg ggaagaacat cttcttgcag
6481 atgtacgaag aaaagctcaa atccagaatc actatcaagg gaaaggagat gagttactat
6541 cagttgctag aatcagaggt gcagaactac aagaacttca tcctgaccgg cgagacatac
6601 aagccgtaca agtactacta gcgtacgtta actttaagaa ggagaactta agatgtatgt
6661 cattctagtt tacgacattc accagaagcg agtaggtaag gctctaaaga tttgtaggaa
6721 gtacctgatc cacatccaga agtcagtttt cgaaggcaac ataacggaga gtaagctaaa
6781 ggccttaaaa gaggagctgg ggcacctgat cgacactcag atggactcgg taattatcta
6841 ccacctggac tccgtgaagt acactaagaa ggagcagatc ggcatcgtcc aatcgacatc
6901 caatgtcatt tgactcgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
6961 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
7021 ggttttttgc tgaaaggagg aactatatcc ggataccagg tgcggggaag cttatgttcc
7081 atagcaaaaa gtcggtcagt ctcgtggctg aaatcatgag ttccacaaaa tggctgaaat
7141 tcaaggaaaa tcaggaatct cagaaaaacg atcgaccgac tttttcgata aaatggttgc
7201 aaaaatgaga aaaatctgat ttaatagaat ctgaaaacag cggaaatgct gttgtcgtac
7261 tttacctaaa aggaattgaa acgtcccgc
//
