LOCUS       Exported                6516 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pcDNA3-ApoE1, complete sequence.
ACCESSION   KY924485
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6516)
  AUTHORS   Li Z, Li Y, Bi Y, Zhang H, Yao Y, Li Q, Cun W, Dong S.
  TITLE     Extracellular Interactions between Hepatitis C Virus and Secreted 
            Apolipoprotein E
  JOURNAL   J. Virol. (2017) In press
REFERENCE   2  (bases 1 to 6516)
  AUTHORS   Li Z, Li Y, Bi Y, Zhang H, Yao Y, Li Q, Cun W, Dong S.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-APR-2017) The Third Department of Biological Products,
            Institute of Medical Biology, Chinese Academy of Medical Sciences & 
            Peking Union Medical College, 935 Jiaoling Road, Kunming, Yunnan 
            650118, China
REFERENCE   3  (bases 1 to 6516)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6516
                     /organism="Cloning vector pcDNA3-ApoE1"
                     /mol_type="other DNA"
                     /label=derived from Homo sapiens
                     /note="derived from Homo sapiens"
                     /db_xref="taxon:1986172"
     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"
     CDS             958..1911
                     /codon_start=1
                     /gene="ApoE1"
                     /product="ApoE1"
                     /label=ApoE1
                     /note="glutamic acid substituted for lysine at residue 146"
                     /protein_id="ARQ79460.1"
                     /translation="MKVLWAALLVTFLAGCQAKVEQAVETEPEPELRQQTEWQSGQRWE
                     LALGRFWDYLRWVQTLSEQVQEELLSSQVTQELRALMDETMKELKAYKSELEEQLTPVA
                     EETRARLSKELQAAQARLGADMEDVCGRLVQYRGEVQAMLGQSTEELRVRLASHLRKLR
                     ERLLRDADDLQKRLAVYQAGAREGAERGLSAIRERLGPLVEQGRVRAATVGSLAGQPLQ
                     ERAQAWGERLRARMEEMGSRTRDRLDEVKEQVAEVRAKLEEQAQQIRLQAEAFQARLKS
                     WFEPLVEDMQRQWAGLVEKVQAAVGTSAAPVPSDNH"
     gene            958..1911
                     /gene="ApoE1"
                     /label=ApoE1
     polyA_signal    2116..2340
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2386..2814
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2828..3157
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3008..3143
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3224..4018
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4192..4313
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4362..4378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4386..4402
                     /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(4410..4440)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4455..4476
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4764..5349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5520..6380)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6381..6485)
                     /gene="bla"
                     /label=AmpR promoter