| 质粒名称: | pLBH545_Tet-Cas14a1_Locus |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 112501 |
| 存储实验室: | Jennifer Doudna |
|
载体骨架基本信息 |
|
| 载体名称: | pTet |
| 完整质粒大小: | 4607bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氯霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Cas14a1/Un1Cas12f1 |
| 插入大小: | 2116 bp |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | gtgccgatcaacgtAtcattttcg |
| 3’测序引物 | acgcagaaaggcccacccgaag |
|
文献引用方法 |
|
| 材料和方法部分: | pLBH545_Tet-Cas14a1_Locus was a gift from Jennifer Doudna (Addgene plasmid # 112501 ; http://n2t.net/addgene:112501 ; RRID:Addgene_112501) |
| 参考文献部分: | Programmed DNA destruction by miniature CRISPR-Cas14 enzymes. Harrington LB, Burstein D, Chen JS, Paez-Espino D, Ma E, Witte IP, Cofsky JC, Kyrpides NC, Banfield JF, Doudna JA. Science. 2018 Oct 18. pii: science.aav4294. doi: 10.1126/science.aav4294. 10.1126/science.aav4294 PubMed 30337455 |
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 pLBH545_Tet-Cas14a1_Locus 4607 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses Cas14a1 locus (including crRNA and tracrRNA) under control
of a tetracycline inducible promoter.
ACCESSION .
VERSION .
KEYWORDS pLBH545_Tet-Cas14a1_Locus
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4607)
AUTHORS Harrington LB, Burstein D, Chen JS, Paez-Espino D, Ma E, Witte IP,
Cofsky JC, Kyrpides NC, Banfield JF, Doudna JA
TITLE Programmed DNA destruction by miniature CRISPR-Cas14 enzymes.
JOURNAL Science. 2018 Oct 18. pii: science.aav4294. doi:
10.1126/science.aav4294.
PUBMED 30337455
REFERENCE 2 (bases 1 to 4607)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4607
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 42..97
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 78..96
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
RBS 115..126
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
terminator 2252..2279
/label=T7Te terminator
/note="phage T7 early transcription terminator"
primer_bind complement(2307..2324)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2441..2986)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(2521..2540)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 3100..3194
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(3215..3874)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 3789..3808
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(3875..3977)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS complement(4007..23)
/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 4538..4557
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
ORIGIN
1 ttacttttat ctaatctaga catcattaat tcctaatttt tgttgacact ctatcgttga
61 tagagttatt ttaccactcc ctatcagtga tagagaaaag aattcaaaag atctaaagag
121 gagaaaggat ctatggccaa aaacaccatt accaaaacac tgaaactgcg tattgtgcgt
181 ccgtataata gcgcagaagt ggaaaaaatt gttgccgacg aaaaaaacaa ccgcgaaaaa
241 atcgcactgg aaaagaacaa agacaaagtg aaagaagcct gcagcaaaca tctgaaagtt
301 gcagcatatt gtaccacaca ggttgaacgt aatgcatgcc tgttttgtaa agcacgtaaa
361 ctggatgaca aattctacca aaaactgcgt ggtcagtttc cggatgcagt tttttggcaa
421 gaaatcagcg aaatttttcg ccagctgcag aaacaggcag cagaaatcta taatcagagc
481 ctgatcgaac tgtactacga gatttttatc aaaggcaaag gtattgcaaa tgccagcagc
541 gttgaacatt atctgagtga tgtttgttat acccgtgcag cagaactgtt taaaaacgca
601 gcaattgcaa gcggtctgcg tagcaaaatc aaaagcaatt ttcgtctgaa agaactgaaa
661 aacatgaaaa gtggtctgcc gaccaccaaa agcgataatt ttccgattcc gctggttaaa
721 cagaaaggtg gtcagtatac cggttttgaa attagcaatc ataatagcga cttcatcatc
781 aagattccgt ttggtcgttg gcaggtcaaa aaagagattg ataaatatcg tccgtgggag
841 aaatttgact ttgaacaggt tcagaaaagc ccgaaaccga ttagcctgct gctgagcacc
901 cagcgtcgta aacgtaataa aggttggagc aaagatgaag gcaccgaagc cgaaatcaaa
961 aaagttatga atggcgatta tcagaccagc tacattgaag ttaaacgtgg cagcaaaatc
1021 ggtgaaaaaa gcgcatggat gctgaatctg agcattgatg ttccgaaaat tgataaaggt
1081 gtggatccga gcattattgg tggtattgat gttggtgtta aatcaccgct ggtttgcgca
1141 attaacaatg catttagccg ttatagcatc agcgataacg acctgtttca cttcaacaag
1201 aaaatgtttg cacgtcgtcg tatcctgctg aaaaaaaacc gtcataaacg tgcaggtcat
1261 ggtgcaaaaa acaaactgaa accgatcacc attctgaccg aaaaaagtga acgttttcgc
1321 aaaaagctga ttgaacgttg ggcatgtgaa atcgcggatt tcttcattaa aaacaaagtt
1381 ggcaccgtgc agatggaaaa tctggaaagc atgaaacgta aagaggacag ctattttaac
1441 attcgcctgc gtggcttttg gccgtatgca gaaatgcaga acaaaatcga attcaaactg
1501 aagcagtatg gcatcgaaat tcgtaaagtt gcaccgaata ataccagcaa aacctgtagc
1561 aaatgtggcc atctgaacaa ctatttcaac ttcgagtacc gcaagaaaaa caaattcccg
1621 cactttaaat gcgaaaaatg caacttcaaa gaaaacgccg attataatgc agccctgaat
1681 atttcaaacc cgaaactgaa aagcaccaaa gaggaaccgt aaatatttat actttattat
1741 ccttcattga caaaaatgag aatgttatcc cagataacat ttgatgtaca cagattcaca
1801 cttcactgat aaagtggaga accgcttcac caaaagctgt cccttagggg attagaactt
1861 gagtgaaggt gggctgcttg catcagccta atgtcgagaa gtgctttctt cggaaagtaa
1921 ccctcgaaac aaattcattt ttcctctcca attctgcaca aaaaaaggtg agtccttata
1981 aaccggcgtg cagaacgccg gctcaccttt tttcttcatt cgattttatg cttaaaagcc
2041 gtaaaaacgc ggaattcggc gccgttgcag aacccgaata gacgaatgaa ggaatgcaac
2101 taccttacac cgcttgcgaa agttgtcaga agataatctt gcagaacccg aatagacgaa
2161 tgaaggaatg caatcttgac agagcccgat tgcgttatct ccaggagaaa catataaaag
2221 catcaaccgc tgatcggact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
2281 ttatacctag ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
2341 ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag
2401 atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc
2461 ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat
2521 aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc
2581 ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc
2641 agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc
2701 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa
2761 gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc
2821 cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc
2881 tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt
2941 ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat
3001 taatcagata aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga
3061 agtcagcccc atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc
3121 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
3181 tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc
3241 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
3301 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
3361 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
3421 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
3481 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
3541 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
3601 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt
3661 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
3721 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
3781 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
3841 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
3901 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
3961 ctcttacgtg ccgatcaacg tctcattttc gccagatatc gacgtcttaa gacccacttt
4021 cacatttaag ttgtttttct aatccgcata tgatcaattc aaggccgaat aagaaggctg
4081 gctctgcacc ttggtgatca aataattcga tagcttgtcg taataatggc ggcatactat
4141 cagtagtagg tgtttccctt tcttctttag cgacttgatg ctcttgatct tccaatacgc
4201 aacctaaagt aaaatgcccc acagcgctga gtgcatataa tgcattctct agtgaaaaac
4261 cttgttggca taaaaaggct aattgatttt cgagagtttc atactgtttt tctgtaggcc
4321 gtgtacctaa atgtactttt gctccatcgc gatgacttag taaagcacat ctaaaacttt
4381 tagcgttatt acgtaaaaaa tcttgccagc tttccccttc taaagggcaa aagtgagtat
4441 ggtgcctatc taacatctca atggctaagg cgtcgagcaa agcccgctta ttttttacat
4501 gccaatacaa tgtaggctgc tctacaccta gcttctgggc gagtttacgg gttgttaaac
4561 cttcgattcc gacctcatta agcagctcta atgcgctgtt aatcact
//
