Adenoviral pOTENT-1 shuttle vector with kanamycin selection and CMV promoter and C-terminal FLAG and His tags.
| 载体基本信息 | |||
| 载体名称 | pOTENT-1 | 载体抗性 | Kanamycin |
|---|---|---|---|
| 载体长度 | 7509 bp | 载体类型 | Mammalian Expression Vectors |
| 筛选标记 | Puromycin | 拷贝数 | Low copy number |
| 启动子 | CMV | 克隆方法 | Enzyme Cut |
| 5引物 | T7 promoter | 3引物 | pOT-R |
| 载体标签 | FLAG/His | 表达方法 | Constiutive, Stable / Transient |
pOTENT-1 载体载体图谱
pOTENT-1 载体载体序列
LOCUS Exported 7509 bp ds-DNA circular SYN 08-SEP-2020
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pOTENT-1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7509)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..7509
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 21..123
/note="ITR"
/note="inverted terminal repeat of human adenovirus
serotype 5"
misc_signal 211..361
/note="Ad5 Psi"
/note="packaging signal for adenovirus serotype 5"
enhancer 419..722
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 723..926
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 954..972
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1073..1096
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/note="FLAG"
/translation="DYKDDDDK"
CDS 1106..1123
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
polyA_signal 1294..1415
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter 2342..2671
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2522..2657
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2729..3328
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="PuroR"
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
polyA_signal 3329..3553
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
repeat_region 4464..4566
/note="ITR"
/note="inverted terminal repeat of human adenovirus
serotype 5"
rep_origin complement(4838..5426)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 6258..7052
/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(7100..7480)
/direction=LEFT
/note="M13 ori"
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
