LOCUS       Exported                7478 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCL0161, complete sequence.
ACCESSION   KC218923
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7478)
  AUTHORS   Bayne AC, Boltz D, Owen C, Betz Y, Maia G, Azadi P, Archer-Hartmann 
            S, Zirkle R, Lippmeier JC.
  TITLE     Vaccination against Influenza with Recombinant Hemagglutinin 
            Expressed by Schizochytrium sp. Confers Protective Immunity
  JOURNAL   PLoS ONE 8 (4), E61790 (2013)
  PUBMED    23626728
REFERENCE   2  (bases 1 to 7478)
  AUTHORS   Bayne A-C.V., Lippmeier JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-NOV-2012) Molecular Biology, DSM Nutritional Products,
            6480 Dobbin Rd., Columbia, MD 21045, USA
REFERENCE   3  (bases 1 to 7478)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7478
                     /organism="Expression vector pCL0161"
                     /mol_type="other DNA"
                     /db_xref="taxon:1325484"
     regulatory      1..1063
                     /regulatory_class="promoter"
                     /note="elongation factor 1 promoter"
     CDS             1070..2767
                     /codon_start=1
                     /gene="HA"
                     /product="hemagglutinin"
                     /label=HA
                     /note="codon-optimized for expression in Schizochytrium sp.
                     ATCC 20888"
                     /protein_id="AGK36287.1"
                     /translation="MKAKLLVLLCTFTATYADTICIGYHANNSTDTVDTVLEKNVTVTH
                     SVNLLEDSHNGKLCLLKGIAPLQLGNCSVAGWILGNPECELLISKESWSYIVETPNPEN
                     GTCYPGYFADYEELREQLSSVSSFERFEIFPKESSWPNHTVTGVSASCSHNGKSSFYRN
                     LLWLTGKNGLYPNLSKSYVNNKEKEVLVLWGVHHPPNIGNQRALYHTENAYVSVVSSHY
                     SRRFTPEIAKRPKVRDQEGRINYYWTLLEPGDTIIFEANGNLIAPWYAFALSRGFGSGI
                     ITSNAPMDECDAKCQTPQGAINSSLPFQNVHPVTIGECPKYVRSAKLRMVTGLRNIPSI
                     QSRGLFGAIAGFIEGGWTGMVDGWYGYHHQNEQGSGYAADQKSTQNAINGITNKVNSVI
                     EKMNTQFTAVGKEFNKLERRMENLNKKVDDGFLDIWTYNAELLVLLENERTLDFHDSNV
                     KNLYEKVKSQLKNNAKEIGNGCFEFYHKCNNECMESVKNGTYDYPKYSEESKLNREKID
                     GVKLESMGVYQILAIYSTVASSLVLLVSLGAISFWMCSNGSLQCRICI"
     gene            1070..2767
                     /gene="HA"
                     /label=HA
                     /note="derived from Influenza A virus (A/New 
                     Caledonia/20/1999(H1N1))"
     regulatory      2787..3426
                     /regulatory_class="terminator"
                     /note="Pfa 3 terminator"
     polyA_signal    3569..3690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3952..4540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4711..5571)
                     /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(5572..5676)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      5961..6409
                     /regulatory_class="promoter"
                     /note="alpha-tubulin promoter"
     CDS             6410..7204
                     /codon_start=1
                     /gene="npt"
                     /product="neomycin phosphotransferase"
                     /label=npt
                     /note="selectable marker"
                     /protein_id="AGK36288.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            6410..7204
                     /gene="npt"
                     /label=npt
     regulatory      7205..7478
                     /regulatory_class="terminator"
                     /note="SV40 terminator"
     polyA_signal    7334..7455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"