pCI- Neomycin Plasmid

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  • Model: PVT1033
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pCI-Neomycin

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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

 

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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