| 载体基本信息 |
| 载体名称 |
pY001 (pFnCpf1_full) |
载体抗性 |
Chloramphenicol |
| 载体长度 |
10653 bp |
载体类型 |
Bacterial Expression, CRISPR |
| 拷贝数 |
Low Copy |
pY001 (pFnCpf1_full) 载体载体图谱
pY001 (pFnCpf1_full) 载体载体序列
LOCUS Exported 10653 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses FnCpf1 locus including Cas genes and spacers 1-4 of CRISPR
array..
ACCESSION .
VERSION .
KEYWORDS pY001 (pFnCpf1_full)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10653)
AUTHORS Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS,
Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A, Koonin
EV, Zhang F
TITLE Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas
System.
JOURNAL Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi:
10.1016/j.cell.2015.09.038.
PUBMED 26422227
REFERENCE 2 (bases 1 to 10653)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2015
Sep 23. pii: S0092-8674(15)01200-3. doi:
10.1016/j.cell.2015.09.038."
FEATURES Location/Qualifiers
source 1..10653
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 134..153
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(220..322)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(714..731)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(848..1393)
/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(928..947)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS 7989..9179
/codon_start=1
/gene="tet"
/product="tetracycline efflux protein"
/label=TcR
/note="confers resistance to tetracycline"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
primer_bind complement(8341..8360)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
CDS complement(join(10213..10653,1..219))
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"