LOCUS       Exported                7108 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pSG58, complete sequence.
ACCESSION   KM288848
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7108)
  AUTHORS   Goldfless SJ, Wagner JC, Niles JC.
  TITLE     Versatile control of Plasmodium falciparum gene expression with an 
            inducible protein-RNA interaction
  JOURNAL   Nat Commun 5, 5329 (2014)
  PUBMED    25370483
REFERENCE   2  (bases 1 to 7108)
  AUTHORS   Goldfless SJ, Niles JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-AUG-2014) Biological Engineering, MIT, 77 
            Massachusetts Avenue, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 7108)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7108
                     /organism="Expression vector pSG58"
                     /mol_type="other DNA"
                     /note="TetR expression vector for integration in Plasmodium
                     falciparum"
                     /db_xref="taxon:1569330"
     promoter        complement(7..25)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(43..59)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    67..83
                     /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)."
     promoter        complement(91..121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    136..157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      445..1033
     CDS             complement(1204..2064)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AIW62879.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2065..2169)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(2065..2134)
                     /regulatory_class="promoter"
                     /note="bla promoter for E. coli"
     primer_bind     2643..2659
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2666..2683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_recomb     2689..2738
                     /label=Bxb1 attP
                     /note="Bxb1 attP"
     misc_recomb     2745..2795
                     /label=Bxb1 attP
                     /note="Bxb1 attP"
     regulatory      complement(2809..3405)
                     /regulatory_class="terminator"
                     /note="derived from P. falciparum HRPII 3 UTR"
     CDS             complement(3411..3812)
                     /codon_start=1
                     /product="blasticidin S deaminase"
                     /label=blasticidin S deaminase
                     /note="blasticidin deaminase selectable marker"
                     /protein_id="AIW62881.1"
                     /translation="MHAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFT
                     GVNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPG
                     IKAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      complement(3824..4418)
                     /regulatory_class="promoter"
                     /note="contains promoter and 5 UTR from P. chabaudi 
                     dihydrofolate reductase-thymidylate synthase gene"
     regulatory      4501..5512
                     /regulatory_class="promoter"
                     /note="contains promoter and 5 UTR from P. falciparum 
                     calmodulin gene"
     CDS             5517..6227
                     /codon_start=1
                     /product="T7 tag-TetR(B)-Hexa-His tag"
                     /label=T7 tag-TetR
                     /protein_id="AIW62880.1"
                     /translation="MASMTGGQQMGRGSEFMSRLDKSKVINSALELLNEVGIEGLTTRK
                     LAQKLGVEQPTLYWHVKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRC
                     ALLSHRDGAKVHLGTRPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLE
                     DQEHQVAKEERETPTTDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG
                     SKLAAALEHHHHHH"
     misc_feature    5517..5567
                     /label=Region: T7 epitope tag
                     /note="Region: T7 epitope tag"
     CDS             5517..5549
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             5565..6185
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     misc_feature    5574..6185
                     /note="Region: TetR protein region; tetR(B)"
     misc_feature    6186..6206
                     /label=Region: linker
                     /note="Region: linker"
     misc_feature    6207..6224
                     /note="Region: Hxa-His tag; hexahistidine affinity tag"
     CDS             6207..6224
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      6233..7069
                     /regulatory_class="terminator"
                     /note="derived from P. falciparum hsp86 3 UTR"