LOCUS       Exported                8685 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p-REX2mDHFR-int, complete sequence.
ACCESSION   KU998258
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8685)
  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 8685)
  AUTHORS   Blancke Soares A, 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 8685)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8685
                     /organism="Cloning vector p-REX2mDHFR-int"
                     /mol_type="other DNA"
                     /db_xref="taxon:1853255"
     regulatory      36..1499
                     /regulatory_class="promoter"
                     /note="Pfcrt 5 region"
     misc_feature    1506..1787
                     /note="similar to REX2; P. falciparum ring exported protein
                     2 (PF3D7_0936000)"
     misc_feature    1794..2354
                     /note="similar to mDHFR; mouse dihydrofolate reductase"
     CDS             1794..2354
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     CDS             2361..3077
                     /codon_start=1
                     /product="GFP"
                     /label=GFP
                     /note="green fluorescent protein"
                     /protein_id="ANH10904.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      3089..3541
                     /regulatory_class="terminator"
                     /note="P. berghei DT 3 region"
     regulatory      complement(3553..3898)
                     /regulatory_class="promoter"
                     /note="PfHOP 5 region"
     regulatory      3899..4389
                     /regulatory_class="promoter"
                     /note="PfCAM 5 region"
     misc_feature    4396..4956
                     /note="similar to hDHFR; human dihydrofolate reductase"
     misc_feature    4963..5019
                     /label=T2A sequence
                     /note="T2A sequence"
     CDS             4966..5019
                     /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             5026..5424
                     /codon_start=1
                     /product="BSD"
                     /label=BSD
                     /note="blasticidin-S deaminase"
                     /protein_id="ANH10905.1"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5429..5998
                     /regulatory_class="terminator"
                     /note="PfhrpII 3 region"
     promoter        complement(6003..6020)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    6004..8685
                     /label=pGEM backbone
                     /note="pGEM backbone"
     primer_bind     complement(6027..6043)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6517..6621
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6622..7482
                     /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      7653..8241
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8529..8550
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8565..8595
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8603..8619
                     /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     8627..8643
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8661..8679
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"