The vector pShuttle-CMV contains a multiple cloning site sandwiched between the CMV promoter and the SV40 polyadenylation signal and is suitable for insertion of a large cDNA (up to 6.6 kb). pShuttle contains only a multiple cloning site. This allows for the insertion of an entire expression cassette, including specialized promoters and termination signals (up to 7.5 kb). The regions indicated as arms are the stretches of sequence homology with pAdEasy-1 where the homologous recombination occurs. The R-ITR and L-ITR regions are short inverted terminal repeats (Left and Right) which have a role in replication of the viral DNA.
| 载体基本信息 | |||
| 载体名称 | pShuttle-CMV | 载体抗性 | Kanamycin |
|---|---|---|---|
| 载体长度 | 7469 bp | 载体类型 | Adenovirus Systems |
| 拷贝数 | Low copy number | 启动子 | CMV |
| 克隆方法 | Enzyme digestion and ligation | 5引物 | pShuttle-CMV-F: GGTCTATATAAGCAGAGCTG |
| 3引物 | pShuttle-CMV-R: GTGGTATGGCTGATTATGATCAG | ||
pShuttle-CMV 载体载体图谱
pShuttle-CMV 载体载体序列
LOCUS Exported 7469 bp ds-DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7469)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..7469
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..103
/note="ITR"
/note="inverted terminal repeat of human adenovirus
serotype 5"
misc_signal 191..341
/note="Ad5 Psi"
/note="packaging signal for adenovirus serotype 5"
enhancer 399..702
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 703..906
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
polyA_signal 1117..1238
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
repeat_region 4429..4531
/note="ITR"
/note="inverted terminal repeat of human adenovirus
serotype 5"
rep_origin complement(4791..5379)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 6211..7005
/codon_start=1
/gene="aph(3)-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
rep_origin complement(7053..7432)
/direction=LEFT
/note="M13 ori"
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
