LOCUS       Exported                8184 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBSD-GFP-INT-attP, complete sequence.
ACCESSION   DQ813652
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8184)
  AUTHORS   Nkrumah LJ, Muhle RA, Moura PA, Ghosh P, Hatfull GF, Jacobs WR Jr., 
            Fidock DA.
  TITLE     Efficient site-specific integration in Plasmodium falciparum 
            chromosomes mediated by mycobacteriophage Bxb1 integrase
  JOURNAL   Nat. Methods 3 (8), 615-621 (2006)
  PUBMED    16862136
REFERENCE   2  (bases 1 to 8184)
  AUTHORS   Nkrumah LJ, Moura PA, Fidock DA.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2006) Microbiology and Immunology, Albert Einstein
            College of Medicine, 1300 Morris Park, Bronx, NY 10461, USA
REFERENCE   3  (bases 1 to 8184)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8184
                     /organism="Cloning vector pBSD-GFP-INT-attP"
                     /mol_type="other DNA"
                     /db_xref="taxon:393298"
     regulatory      complement(37..865)
                     /regulatory_class="terminator"
                     /note="derived from hsp86 3 UTR"
     CDS             complement(872..2020)
                     /codon_start=1
                     /gene="bsd-FLAG-gfp"
                     /product="BSD-FLAG-GFP"
                     /label=bsd-FLAG-gfp
                     /note="blasticidin S deaminase-FLAG tag-GFP Mut2 fusion; 
                     selectable marker"
                     /protein_id="ABG81364.1"
                     /translation="MHAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFT
                     GVNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPG
                     IKAIVKDSDGQPTAVGIRELLPSGYVWEGTSDYKDDDDKRSSKGEELFTGVVPILVELD
                     GDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHM
                     KQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNIL
                     GHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLP
                     DNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK"
     gene            complement(872..2020)
                     /gene="bsd-FLAG-gfp"
                     /label=bsd-FLAG-gfp
     misc_feature    complement(872..1585)
                     /gene="bsd-FLAG-gfp"
                     /label=GFP
                     /note="GFP"
     CDS             complement(872..1585)
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     misc_feature    complement(1592..1615)
                     /gene="bsd-FLAG-gfp"
                     /label=FLAG tag
                     /note="FLAG tag"
     CDS             complement(1592..1615)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    complement(1622..2020)
                     /gene="bsd-FLAG-gfp"
                     /label=BSD
                     /note="BSD"
     CDS             complement(1622..2020)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MHAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFT
                     GVNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPG
                     IKAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    complement(2036..2659)
                     /label=contains promoter and 5 UTR from calmodulin gene
                     /note="contains promoter and 5 UTR from calmodulin gene"
     misc_feature    2666..3264
                     /label=contains promoter and 5 UTR from PcDT gene
                     /note="contains promoter and 5 UTR from PcDT gene"
     CDS             3281..4831
                     /codon_start=1
                     /product="integrase-myc"
                     /label=integrase-myc
                     /note="myc-tagged integrase; derived from mycobacteriophage
                     Bxb1"
                     /protein_id="ABG81365.1"
                     /translation="MRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLD
                     VSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLV
                     VSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGY
                     LPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQG
                     REPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAE
                     LVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTV
                     AMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAG
                     SPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSM
                     NVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMSRAAALEEQKLISEEDL"
     CDS             4799..4828
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      4838..5428
                     /regulatory_class="terminator"
                     /note="derived from hrp2 3 UTR"
     misc_recomb     5447..5496
                     /label=attP
                     /note="attP"
     promoter        complement(5502..5519)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5526..5542)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6016..6120
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6121..6981
                     /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      7152..7740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8028..8049
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8064..8094
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8102..8118
                     /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     8126..8142
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8160..8178
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"