pCDNA3.1-3*HA-N
PVT10681 2ug
pCDNA3.1-3*HA-N Informaiton
Promoter: CMV promoter
Replicon: pUC ori, F1 ori
Terminator: BGH poly (A) signal
Plasmid classification: lactation serial plasmid; lactation expression plasmid; pCDNA series plasmid.
Plasmid size: 5541bp
Plasmid tagging: N-3 * HA
Prokaryotic resistance: ampicillin Amp (100 ug/ml)
Screening marker: neomycin Neo/G418
Cloning strains: E. coli DH5 alpha
Culture conditions: 37 C, aerobic, LB
Expression host: mammalian cells such as 293T
Culture conditions: 37 C, 5%CO2
Induction mode: no need to induce
5'sequencing primers: pCDNA3.1-F (CTAGAGAACCCACTGCTTAC)
3'sequencing primers: pCDNA3.1-R (TAGAAGGCACAGTCGAGG)
pCDNA3.1-3*HA-N Description
pCDNA3.1-3×HA-N vector is designed for high-level, constitutive expression in a variety of mammalian cell lines. It contains a Geneticin selectable marker and a forward-orientation multiple cloning site.High-level stable and non-replicative transient expression can be carried out in most mammalian cells. The vectors contain the following elements: Human cytomegalovirus immediate-early (CMV) promoter for high-level expression in a wide range of mammalian cells Neomycin resistance gene for selection of stable cell lines. There are three HA tags on the N side.Episomal replication in cells lines that are latently infected with SV40 or that express the SV40 large T antigen.
pCDNA3.1-3*HA-N Cloning site
pCDNA3.1-3*HA-N Sequence
LOCUS Exported 5541 bp ds-DNA circular SYN 18-JUL-2016
DEFINITION synthetic circular DNA
KEYWORDS pCDNA3.1-3×HA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5541)
TITLE Direct Submission
JOURNAL Exported Monday, July 18, 2016
FEATURES Location/Qualifiers
source 1..5541
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 235..614
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 928..954
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/note="HA"
/translation="YPYDVPDYA"
CDS 958..984
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/note="HA"
/translation="YPYDVPDYA"
CDS 988..1014
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/note="HA"
/translation="YPYDVPDYA"
polyA_signal 1141..1365
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 1411..1839
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1853..2182
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2033..2168
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2249..3043
/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 3217..3338
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(3387..3403)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3411..3427
/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(3435..3465)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3480..3501
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3789..4374)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4545..5405)
/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(5406..5510)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 gtttaaactt aagccaccat ggcggcgtac ccatacgacg tacctgatta tgcaacttat
961 ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta cgcatcagcg
1021 gaaaagcttg gtaccgagct cggatccact agtccagtgt ggtggaattc tgcagatatc
1081 cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga
1141 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
1201 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
1261 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
1321 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
1381 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
1441 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
1501 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
1561 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
1621 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
1681 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
1741 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
1801 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
1861 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
1921 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
1981 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
2041 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
2101 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
2161 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
2221 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
2281 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
2341 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
2401 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
2461 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
2521 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
2581 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
2641 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
2701 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
2761 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
2821 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
2881 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
2941 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
3001 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
3061 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
3121 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
3181 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
3241 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
3301 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
3361 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
3421 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
3481 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
3541 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
3601 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
3661 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
3721 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
3781 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
3841 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
3901 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
3961 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
4021 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
4081 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
4141 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
4201 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
4261 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
4321 agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
4381 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
4441 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
4501 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
4561 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
4621 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
4681 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
4741 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
4801 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
4861 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
4921 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
4981 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
5041 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
5101 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
5161 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
5221 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
5281 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
5341 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
5401 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
5461 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
5521 cgaaaagtgc cacctgacgt c
//
Caution:
Product is for research use only!
