LOCUS       Exported                7397 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pcDNA3-N-HA-LIC, complete sequence.
ACCESSION   KT359064
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7397)
  AUTHORS   Savitsky PA.
  TITLE     Mammalian expression vector with N-terminal HA tag
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7397)
  AUTHORS   Savitsky PA.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-AUG-2015) Structural Genomics Consortium, University 
            of Oxford, Old Road Campus Research Building, Roosevelt Drive, 
            Oxford, Oxfordshire OX3 7DQ, UK
REFERENCE   3  (bases 1 to 7397)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7397
                     /organism="Mammalian expression vector pcDNA3-N-HA-LIC"
                     /mol_type="other DNA"
                     /label=derived from Escherichia coli
                     /note="derived from Escherichia coli"
                     /db_xref="taxon:1758149"
     regulatory      232..819
                     /regulatory_class="promoter"
                     /note="CMV promoter"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      863..879
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    901..927
                     /label=HA tag
                     /note="HA tag"
     CDS             901..927
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    937..950
                     /label=LIC5
                     /note="LIC5"
     promoter        964..1409
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             1410..2831
                     /codon_start=1
                     /gene="SacB"
                     /product="levansucrase"
                     /EC_number="2.4.1.10"
                     /label=SacB
                     /note="converts sucrose to a toxic product; allows 
                     selection of recombinant plasmids on plates containing 5% 
                     sucrose"
                     /protein_id="ALO78650.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYKVPEFDSSTIKNISSAKGLDVWDSWPLQNTDGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            1410..2831
                     /gene="SacB"
                     /label=SacB
     misc_feature    2877..2891
                     /label=LIC3
                     /note="LIC3"
     regulatory      2993..3217
                     /regulatory_class="polyA_signal_sequence"
                     /note="bGH polyA"
     polyA_signal    2993..3217
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3263..3691
                     /label=F1 replication origin
                     /note="F1 replication origin"
     regulatory      3696..4066
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        3705..4035
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3886..4021
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4102..4896
                     /codon_start=1
                     /gene="NeoR"
                     /product="aminoglycoside phosphotransferase"
                     /EC_number="2.7.1.95"
                     /label=NeoR
                     /note="catalyzes the addition of phosphate from ATP to the 
                     3-hydroxyl group of kanamycin; confers resistance to 
                     kanamycin in bacteria or geneticin (G418) in eukaryotic 
                     cells"
                     /protein_id="ALO78651.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            4102..4896
                     /gene="NeoR"
                     /label=NeoR
     regulatory      5070..5200
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 polyA"
     polyA_signal    5070..5191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5240..5256)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5264..5280
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(5288..5318)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5333..5354
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      5583..6256
                     /label=pUC replication origin
                     /note="pUC replication origin"
     rep_origin      complement(5642..6230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6401..7261)
                     /codon_start=1
                     /gene="AmpR"
                     /product="beta-lactamase"
                     /EC_number="3.5.2.6"
                     /label=AmpR
                     /protein_id="ALO78652.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSCGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6401..7261)
                     /gene="AmpR"
                     /label=AmpR
                     /note="ampicillin resistance protein"
     promoter        complement(7262..7366)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(7262..7360)
                     /regulatory_class="promoter"
                     /note="bla promoter"