LOCUS       Exported               10163 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTGPI-GFP, complete sequence.
ACCESSION   AY860671
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10163)
  AUTHORS   Meissner M, Krejany E, Gilson PR, de Koning-Ward TF, Soldati D, 
            Crabb BS.
  TITLE     Tetracycline analogue-regulated transgene expression in Plasmodium 
            falciparum blood stages using Toxoplasma gondii transactivators
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 102 (8), 2980-2985 (2005)
  PUBMED    15710888
REFERENCE   2  (bases 1 to 10163)
  AUTHORS   Meissner M, Krejany E, Gilson P, de Koning Ward T, Soldati D, Crabb 
            B.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-DEC-2004) Infection and Immunity, Walter and Eliza 
            Hall Institute of Medical Research, Royal Parade, Melbourne, 
            Victoria 3050, Australia
REFERENCE   3  (bases 1 to 10163)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10163
                     /organism="Expression vector pTGPI-GFP"
                     /lab_host="Plasmodium falciparum"
                     /mol_type="other DNA"
                     /db_xref="taxon:308864"
     regulatory      1..1078
                     /regulatory_class="promoter"
                     /note="calmodulin promoter"
     CDS             1079..1642
                     /codon_start=1
                     /gene="hDHFR"
                     /product="human dihydrofolate reductase"
                     /label=hDHFR
                     /protein_id="AAW55664.1"
                     /translation="MHGSLNCIVAVSQNMGIGKNGDYPWPPLRNEFRYFQRMTTTSSVE
                     GKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQP
                     ELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEY
                     PGVLSDVQEEKGIKYKFEVYEKND"
     gene            1079..1642
                     /gene="hDHFR"
                     /label=hDHFR
     regulatory      1641..2215
                     /regulatory_class="terminator"
                     /note="HRP23 terminator"
     promoter        complement(2219..2236)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2220..4933
                     /label=Region: pGem plasmid backbone
                     /note="Region: pGem plasmid backbone"
     primer_bind     complement(2243..2259)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2733..2837
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2838..3698
                     /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      3869..4457
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4745..4766
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4781..4811
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4819..4835
                     /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     4843..4859
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4877..4895
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4927..4943)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      4934..5239
                     /regulatory_class="promoter"
                     /note="7 x Tet operator sequence"
     protein_bind    4954..5224
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     regulatory      5240..5563
                     /regulatory_class="promoter"
                     /note="minimal calmodulin promoter"
     CDS             5579..6499
                     /codon_start=1
                     /gene="GFP_GPI"
                     /product="green fluorescent protein with N-terminal signal 
                     peptide and C-terminal GPI anchor signal sequence from 
                     merozoite surface protein 1"
                     /label=GFP_GPI
                     /protein_id="AAW55663.1"
                     /translation="MKIIFFLCSFLFFIINTQCVTHEMSKGEELFTGVVPILVELDGDV
                     NGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQH
                     DFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHK
                     LEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKNVEKDEAHVKITKLSDLKAI
                     VFCSSSGFMGLSILLIITLIVFNLF"
     gene            5579..6499
                     /gene="GFP_GPI"
                     /label=GFP_GPI
     CDS             5648..6361
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      complement(7151..8075)
                     /regulatory_class="terminator"
                     /note="Plasmodium berghei dihydrofolate reductase 
                     terminator"
     CDS             complement(8119..8775)
                     /codon_start=1
                     /gene="TATi3"
                     /product="Tet repressor and transactivator fusion protein"
                     /label=TATi3
                     /protein_id="AAW55665.1"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLTLVPNWNNTQPYLGLFF"
     gene            complement(8119..8775)
                     /gene="TATi3"
                     /label=TATi3
     regulatory      complement(8776..9652)
                     /regulatory_class="promoter"
                     /note="Plasmodium falciparum merozoite surface protein 2 
                     promoter"
     repeat_region   9653..10163
                     /label=REP20 subtelomeric repeat region
                     /note="REP20 subtelomeric repeat region"