LOCUS       Exported                8609 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSLI-PfEHD2xFKBP, complete sequence.
ACCESSION   KU998257
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8609)
  AUTHORS   Mesen-Ramirez P, Reinsch F, Blancke Soares A, Bergmann B, Ullrich 
            AK, Tenzer S, Spielmann T.
  TITLE     Stable Translocation Intermediates Jam Global Protein Export in 
            Plasmodium falciparum Parasites and Link the PTEX Component EXP2 
            with Translocation Activity
  JOURNAL   PLoS Pathog. 12 (5), E1005618 (2016)
  PUBMED    27168322
REFERENCE   2  (bases 1 to 8609)
  AUTHORS   Birnbaum J, Flemming S, Spielmann T.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-MAR-2016) Parasitology Section, Bernhard Nocht 
            Institute for Tropical Medicine, Bernhard-Nocht-Str. 74, Hamburg 
            20359, Germany
REFERENCE   3  (bases 1 to 8609)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8609
                     /organism="Cloning vector pSLI-PfEHD2xFKBP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1853254"
     regulatory      1..453
                     /regulatory_class="terminator"
                     /note="P. berghei 3 region"
     regulatory      complement(465..810)
                     /regulatory_class="terminator"
                     /note="Pfhop 5 region"
     regulatory      811..1301
                     /regulatory_class="promoter"
                     /note="PfCAM 5 region"
     CDS             1308..1871
                     /codon_start=1
                     /product="hDHFR"
                     /label=hDHFR
                     /note="human dihydrofolate reductase"
                     /protein_id="ANH10902.1"
                     /translation="MHGSLNCIVAVSQNMGIGKNGDYPWPPLRNEFRYFQRMTTTSSVE
                     GKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQP
                     ELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEY
                     PGVLSDVQEEKGIKYKFEVYEKND"
     regulatory      1874..2443
                     /regulatory_class="terminator"
                     /note="P. falciparum hrpII 3 region"
     promoter        complement(2448..2465)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2449..5130
                     /label=pGEM backbone
                     /note="pGEM backbone"
     primer_bind     complement(2472..2488)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2962..3066
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3067..3927
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4098..4686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4974..4995
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5010..5040
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5048..5064
                     /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)."
     primer_bind     5072..5088
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5106..5124
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    5142..6257
                     /note="similar to PfEHD; P. falciparum Eps 15 homology 
                     domain protein, C-terminal fragment (162-533)"
     misc_feature    6264..6320
                     /label=linker
                     /note="linker"
     misc_feature    6327..6980
                     /note="similar to 2xFKBP; 2 x FKBP (FK506 binding protein)"
     CDS             6330..6650
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             6657..6977
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    6987..7025
                     /label=linker
                     /note="linker"
     misc_feature    7032..7745
                     /note="similar to GFP; green fluorescent protein"
     misc_feature    7752..7808
                     /label=T2A sequence
                     /note="T2A sequence"
     CDS             7755..7808
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7809..8603
                     /codon_start=1
                     /product="APH"
                     /label=APH
                     /note="neomycin phosphotransferase"
                     /protein_id="ANH10903.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"