| 载体基本信息 |
| 载体名称 |
pcDNA6.2/EmGFP-Bsd/V5-DEST |
载体抗性 |
Ampicillin |
| 载体长度 |
8198 bp |
载体类型 |
Gateway Cloning Vectors |
| 载体来源 |
Invitrogen (Life Technologies) |
拷贝数 |
High copy number |
pcDNA6.2/EmGFP-Bsd/V5-DEST 载体载体图谱
pcDNA6.2/EmGFP-Bsd/V5-DEST 载体载体序列
LOCUS Exported 8198 bp ds-DNA circular SYN 18-11-2013
DEFINITION Vivid Colors(TM) Gateway(R) destination vector for expressing
proteins in mammalian cells, with an EmGFP-Bsd fusion as a
transfection marker.
ACCESSION .
VERSION .
KEYWORDS pcDNA6.2 EmGFP-Bsd V5-DEST
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8198)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8198
/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"
protein_bind 911..1035
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR1"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(1464..1769)
/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"
CDS complement(2114..2773)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(2774..2876)
/note="cat promoter"
/note="promoter of the E. coli cat gene"
protein_bind complement(3054..3178)
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR2"
/note="recombination site for the Gateway(R) LR reaction"
CDS 3204..3245
/codon_start=1
/product="epitope tag from simian virus 5"
/note="V5 tag"
/translation="GKPIPNPLLGLDST"
polyA_signal 3329..3377
/note="HSV TK poly(A) signal"
/note="herpesvirus thymidine kinase polyadenylation signal"
rep_origin 3579..4007
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4022..4521
/note="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
promoter 4535..4582
/note="EM7 promoter"
/note="synthetic bacterial promoter "
CDS 4601..5317
/codon_start=1
/product="Emerald GFP"
/note="EmGFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 5318..5713
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/note="BSD"
/note="confers resistance to blasticidin"
/translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
AIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 5871..5992
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(6443..7031)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7202..8062)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPKLNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(8063..8167)
/gene="bla"
/note="AmpR promoter"