pCI-Neomycin Search name
pCI-Neomycin,Plasmid pCI-Neomycin,pCI-Neomycin vector
pCI-Neomycin Informaiton
romoter: CMV IE
Replicator: pUC ori, F1 ori
Terminator: SV40 poly (A) signal
Plasmid classification: mammalian cell carrier; protein overexpression plasmid.
Plasmid size: 5472bp
Prokaryotic resistance: ampicillin Amp
Screening markers: neomycin Neo
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, aerobic, LB
Expression host: mammalian cells
Culture conditions: 37 centigrade, aerobic, LB
Induction mode: no induction, instantaneous expression
5'sequencing primers: CMV-F: 5-CGCAAATGGGCGGTAGGCGTG-3
3'sequencing primers: EBV-R:5-GTGGTTTGTCCAAACTCATC-3
pCI-Neomycin Description
pCI-neo Mammalian Expression Vector (pCI-neo mammalian expression vector) carrying the immediate early enhancer / promoter region with human cytomegalovirus (CMV) to promote the composition of the cloned DNA in mammalian cells. This vector also contains neomycin phosphotransferase gene, a selective marker of mammalian cells. Through G-418 resistance screening, transfected cells can be used for transient or stable expression of pCI-neo vector.
pCI-Neomycin Sequence
LOCUS Exported 5472 bp ds-DNA circular SYN 06-11-2015
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5472)
AUTHORS admin
TITLE Direct Submission
JOURNAL Exported 2015-11-6
FEATURES Location/Qualifiers
source 1..5472
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 138..517
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 518..729
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
intron 890..1022
/note="chimeric intron"
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
promoter 1067..1085
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(1141..1158)
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
polyA_signal 1176..1297
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 1483..1938
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2055..2412
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2263..2398
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2463..3257
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3321..3369
/note="synthetic polyadenylation signal"
promoter 3675..3779
/gene="bla"
/note="AmpR promoter"
CDS 3780..4640
/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"
rep_origin 4811..5399
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
1081 ataggctagc ctcgagaatt cacgcgtggt acctctagag tcgacccggg cggccgcttc
1141 cctttagtga gggttaatgc ttcgagcaga catgataaga tacattgatg agtttggaca
1201 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
1261 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
1321 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
&nbs <
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