载体基本信息 | |||
载体名称 | pcDNA6.2/nGeneBLAzer-DEST | 载体抗性 | Ampicillin |
---|---|---|---|
载体长度 | 7564 bp | 载体类型 | Gateway Cloning Vectors |
载体来源 | Invitrogen (Life Technologies) | 拷贝数 | High copy number |
pcDNA6.2/nGeneBLAzer-DEST 载体载体图谱
pcDNA6.2/nGeneBLAzer-DEST 载体载体序列
LOCUS Exported 7564 bp ds-DNA circular SYN 16-11-2013 DEFINITION Gateway(R) destination vector for in vivo or in vitro detection of mammalian gene expression using an N-terminal beta-lactamase reporter. ACCESSION . VERSION . KEYWORDS pcDNA6.2 nGeneBLAzer-DEST SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7564) AUTHORS Invitrogen (Life Technologies) TITLE Direct Submission FEATURES Location/Qualifiers source 1..7564 /organism="synthetic DNA construct" /lab_host="Mammalian Cells" /mol_type="other DNA" enhancer 235..614 /note="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /note="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" promoter 863..881 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" CDS 911..1702 /codon_start=1 /gene="bla(M)" /product="beta-lactamase lacking the signal sequence" /note="bla(M)" /note="allows cytosolic expression of beta-lactamase" /translation="MDPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMM STFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAIT MSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMA TTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGII AALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW" protein_bind 1712..1836 /gene="mutant version of attR" /bound_moiety="LR Clonase(TM)" /note="attR1" /note="recombination site for the Gateway(R) LR reaction" promoter 1861..1891 /note="lac UV5 promoter" /note="E. coli lac promoter with an ""up"" mutation" CDS 1945..2604 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /note="CmR" /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" CDS 2946..3251 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /note="ccdB" /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" protein_bind complement(3292..3416) /gene="mutant version of attR" /bound_moiety="LR Clonase(TM)" /note="attR2" /note="recombination site for the Gateway(R) LR reaction" CDS 3424..3465 /codon_start=1 /product="epitope tag from simian virus 5" /note="V5 tag" /translation="GKPIPNPLLGLDST" polyA_signal 3549..3597 /note="HSV TK poly(A) signal" /note="herpesvirus thymidine kinase polyadenylation signal" rep_origin 3799..4227 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 4241..4570 /note="SV40 promoter" /note="SV40 enhancer and early promoter" rep_origin 4421..4556 /note="SV40 ori" /note="SV40 origin of replication" promoter 4618..4665 /note="EM7 promoter" /note="synthetic bacterial promoter " CDS 4684..5082 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /note="BSD" /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" polyA_signal 5240..5361 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" rep_origin complement(5812..6397) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6568..7428) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(7429..7533) /gene="bla" /note="AmpR promoter"
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