Basic Vector Information
Adenoviral pENTER shuttle vector with kanamycin selection and CMV promoter and C-terminal FLAG and His tags.
pENTER vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pENTER vector Sequence
LOCUS Exported 7509 bp DNA circular SYN 04-NOV-2023 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7509) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..7509 /mol_type="other DNA" /organism="synthetic DNA construct" repeat_region 21..123 /label=ITR /note="inverted terminal repeat of human adenovirus serotype 5" misc_signal 211..361 /label=Ad5 Psi /note="packaging signal for adenovirus serotype 5" enhancer 419..722 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 723..926 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" promoter 954..972 /label=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" /label=FLAG /translation="DYKDDDDK" CDS 1106..1123 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 1294..1415 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter 2342..2671 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2522..2657 /label=SV40 ori /note="SV40 origin of replication" CDS 2729..3328 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" polyA_signal 3329..3553 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 4464..4566 /label=ITR /note="inverted terminal repeat of human adenovirus serotype 5" rep_origin complement(4838..5426) /direction=LEFT /label=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" /label=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 /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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