LOCUS       Exported                8799 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector AF286077, complete sequence.
ACCESSION   AF286077
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8799)
  AUTHORS   Wang RF, Mullins JI.
  TITLE     Mammalian cell/vaccinia virus expression vectors with increased 
            stability of retroviral sequences in Escherichia coli: production of
            feline immunodeficiency virus envelope protein
  JOURNAL   Gene 153 (2), 197-202 (1995)
  PUBMED    7875588
REFERENCE   2  (bases 1 to 8799)
  AUTHORS   Mullins JI, Hanley T, Kafsack BFC., Rowen L.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUL-2000) Microbiology, University of Washington, HSB 
            K-455, Seattle, Washington 98195, USA
REFERENCE   3  (bases 1 to 8799)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8799
                     /organism="Expression vector AF286077"
                     /mol_type="other DNA"
                     /db_xref="taxon:146704"
     misc_feature    join(1..500,8708..8799)
                     /label=vaccinia thymidine kinase gene cluster 5 end
                     /note="vaccinia thymidine kinase gene cluster 5 end"
     gene            46..500
                     /pseudo
                     /gene="thymidine kinase"
                     /label=thymidine kinase
                     /note="derived from vaccinia; 5 end"
     regulatory      502..864
                     /regulatory_class="promoter"
                     /note="derived from SV40"
     promoter        526..1140
                     /label=SR-alpha promoter
                     /note="hybrid promoter consisting of the SV40 early 
                     promoter plus part of the long terminal repeat of human 
                     T-lymphotrophic virus 1"
     rep_origin      706..841
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      872..1306
                     /regulatory_class="promoter"
                     /note="SR-alpha Hybrid Promoter; formed by HTLV LTR and 
                     SV40 promoter"
     LTR             872..1140
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     intron          1181..1274
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     promoter        1302..1320
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1344..1892
                     /regulatory_class="promoter"
                     /note="derived from EMV"
     misc_feature    1849..1857
                     /label=oligopyrimidine tract
                     /note="oligopyrimidine tract"
     misc_feature    1917..1934
                     /label=polyhistidine tract
                     /note="polyhistidine tract"
     CDS             1917..1934
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    1960..2007
                     /label=similar to tpa 3 end
                     /note="similar to tpa 3 end"
     regulatory      2018..2288
                     /regulatory_class="polyA_signal_sequence"
                     /note="derived from bovine growth hormone"
     polyA_signal    2031..2255
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      2301..2331
                     /regulatory_class="polyA_signal_sequence"
                     /note="synthetic"
     regulatory      2339..2465
                     /regulatory_class="terminator"
                     /note="derived from T7"
     terminator      2396..2443
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      2474..2747
                     /regulatory_class="promoter"
                     /note="derived from P7.5 vaccinia"
     misc_feature    2777..3143
                     /label=murine beta-globin transcriptional regulation unit
                     /note="murine beta-globin transcriptional regulation unit"
     misc_feature    3189..3851
                     /label=DHFR
                     /note="DHFR"
     CDS             3244..3807
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     regulatory      3888..4141
                     /regulatory_class="polyA_signal_sequence"
                     /note="derived from bovine growth hormone"
     polyA_signal    3900..4124
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    4142..4507
                     /label=murine beta-globin transcriptional regulation unit
                     /note="murine beta-globin transcriptional regulation unit"
     gene            4527..5492
                     /gene="neomycin resistance"
                     /label=neomycin resistance
     CDS             4528..5322
                     /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="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      5496..5632
                     /regulatory_class="polyA_signal_sequence"
                     /note="derived from SV40 hormone"
     polyA_signal    5496..5630
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    5638..6265
                     /label=vaccinia thymidine kinase gene cluster 3 end
                     /note="vaccinia thymidine kinase gene cluster 3 end"
     gene            5638..5911
                     /pseudo
                     /gene="thymidine kinase"
                     /label=thymidine kinase
                     /note="derived from vaccinia; 3 end"
     misc_feature    6320..6449
                     /label=similar to pUC19
                     /note="similar to pUC19"
     rep_origin      6453..7047
                     /label=p15A intermediate copy number
                     /note="p15A intermediate copy number"
     rep_origin      6489..7033
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     misc_feature    7050..8192
                     /label=similar to pUC19
                     /note="similar to pUC19"
     CDS             complement(7192..8052)
                     /codon_start=1
                     /product="beta lactamase"
                     /label=beta lactamase
                     /protein_id="AAG45964.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8053..8157)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8183..8638)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     misc_feature    8210..8636
                     /label=f1 packaging origin
                     /note="f1 packaging origin"