p3xFLAG-CMV-14
PVT1046 2ug
p3xFLAG-CMV-14 Information
Promoter: CMV
Replicon: pUC
Terminator: SV40 poly (a) signal, hGH poly (a) signal
Plasmid classification: mammalian cells, protein overexpression vector
Plasmid size: 6310bp
Prokaryotic resistance: Amp
Eukaryotic resistance: G418
Clone strain: DH5a
Culture conditions: 37℃
Expression host: Mammalian cells
Induction mode: transient expression without induction
5 'sequencing primer: cmv-f: cgcaaaatgggcgtgtgtgggtgggtg
3 'sequencing primer: hgh-pa-r: ccagttgtcccaataga
Plasmid host: mammalian cells
Purpose of plasmid: protein expression
Fragment type: ORF
Fragment species: empty bodies
Prokaryotic resistance: Amp
Eukaryotic resistance: G418
p3xFLAG-CMV-14 Description
p3xFLAG-CMV-14 vector is a 6.3 KB carrier derived from the pCMV5 vector, which is used for transient expression in mammalian cells or the stable expression of C terminal with 3xFlag tag fusion protein. Three adjacent Flag epitopes (Asp-Tyr-Lys-Xaa-Xaa-Asp) were encoded by the downstream fragment of the vector. This makes it possible to detect protein expression using M2 Flag antibody.
p3xFLAG-CMV-14 Sequence
LOCUS Exported 6310 bp ds-DNA circular SYN 29-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 12
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6310)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Monday, August 29, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..6310
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 141..157
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
enhancer 318..697
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 698..901
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 993..1058
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/note="3xFLAG"
/translation="DYKDHDGDYKDHDIDYKDDDDK"
polyA_signal 1069..1691
/note="hGH poly(A) signal"
/note="human growth hormone polyadenylation signal"
promoter 1733..2030
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 2084..2878
/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 3530..3664
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(3670..3688)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3702..3718)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3726..3742
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(3750..3780)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3795..3816
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4104..4692)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4863..5723)
/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"
promoter complement(5724..5828)
/gene="bla"
/note="AmpR promoter"
rep_origin 5855..6310
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
901 tcgtttagtg aaccgtcaga attaagcttg cggccgcgaa ttcatcgata gatctgatat
961 cggtaccagt cgactctaga ggatcccggg ctgactacaa agaccatgac ggtgattata
1021 aagatcatga catcgactac aaggatgacg atgacaagta gtgatcccgg gtggcatccc
1081 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
1141 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
1201 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
1261 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
1321 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
1381 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
1441 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
1501 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa
1561 ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg
1621 ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg
1681 cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga
1741 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
1801 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
1861 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
1921 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
1981 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggaattgat
2041 cagcttggga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
2101 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
2161 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
2221 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
2281 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
2341 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
2401 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
2461 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
2521 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
2581 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
2641 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
2701 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
2761 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
2821 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
2881 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
2941 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
3001 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc cgggctcgat
3061 cccctcgcga gttggttcag ctgctgcctg aggctggacg acctcgcgga gttctaccgg
3121 cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct atccgcgcat ccatgccccc
3181 gaactgcagg agtggggagg cacgatggcc gctttggtcg acccggacgg gacgctcctg
3241 cgcctgatac agaacgaatt gcttgcaggc atctcatgag tgtgtcttcc cgttttccgc
3301 ctgaggtcac tgcgtggatg gagcgctggc gcctgctgcg cgacggcgag ctgctcacca
3361 cccactcgcc aagctggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
3421 gccgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
3481 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
3541 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
3601 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
3661 gatcaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg
3721 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
3781 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
3841 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
3901 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
3961 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
4021 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
4081 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
4141 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
4201 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
4261 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
4321 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
4381 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
4441 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
4501 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
4561 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
4621 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
4681 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
4741 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
4801 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
4861 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
4921 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
4981 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
5041 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
5101 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
5161 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
5221 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
5281 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
5341 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
5401 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
5461 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
5521 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
5581 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
5641 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
5701 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
5761 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
5821 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
5881 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
5941 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
6001 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
6061 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
6121 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
6181 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
6241 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
6301 cttacaattt
//
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Product is for research use only!
