LOCUS       Exported                4445 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBSK-mBC4-Sense-GUS-Antisense, complete sequence.
ACCESSION   HQ680380
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4445)
  AUTHORS   Narwade AV, Chakrabarty PK, Kalbande BB, Chavhan RL, Warade JG, 
            Sable SV, Nandeshwar SB.
  TITLE     Inverted repeat construct for RNA interference-mediated targeting of
            beta-C4 protein gene of Cotton Leaf Curl Virus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4445)
  AUTHORS   Narwade AV, Chakrabarty PK, Kalbande BB, Chavhan RL, Warade JG, 
            Sable SV, Nandeshwar SB.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-DEC-2010) Division of Crop Improvement, Central 
            Institute for Cotton Research, Post Bag No. 02, Shankar Nagar P.O., 
            Nagpur, Maharashtra 440010, India
REFERENCE   3  (bases 1 to 4445)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4445
                     /organism="Cloning vector pBSK-mBC4-Sense-GUS-Antisense"
                     /mol_type="other DNA"
                     /label=modified inverted repeat construct
                     /note="modified inverted repeat construct"
                     /db_xref="taxon:1076682"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      35..441
                     /label=F1(+) origin of replication
                     /note="F1(+) origin of replication"
     misc_feature    complement(460..2300)
                     /label=beta-galactosidase alpha (lacZ) fragment
                     /note="beta-galactosidase alpha (lacZ) fragment"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    653..2244
                     /note="MCS; multiple cloning site"
     misc_feature    659..991
                     /note="mBC4-sense strand; modified beta-C4 protein fragment
                     of Cotton Leaf Curl Virus isolate Sirsa (Haryana) segment 
                     beta-DNA deposite in GenBank Accession NumberHM146308; 
                     cloned in sense orientation"
     misc_feature    998..1875
                     /note="beta-glucuronidase; Escherichia coli GusA (N358Q) 
                     fragment from pCAMBIA-1302 was PCR amplified with 
                     engineered restriction sites and cloned in EcoRI and BamHI 
                     sites of the MCS of lacZ in pBluscriptII SK(+) (X52328.1)"
     misc_feature    1882..2214
                     /note="mBC4-antisense strand; modified beta-C4 protein 
                     fragment of Cotton Leaf Curl Virus isolate Sirsa (Haryana) 
                     segment beta-DNA deposite in GenBank Accession 
                     NumberHM146308; cloned in antisense orientation"
     promoter        complement(2257..2275)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2296..2312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2301..2422
                     /regulatory_class="promoter"
                     /note="lac promoter"
     protein_bind    2320..2336
                     /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(2344..2374)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2389..2410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2642..3309
                     /label=pUC origin of replication
                     /note="pUC origin of replication"
     rep_origin      complement(2698..3286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3457..4317)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AEM60379.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4318..4422)
                     /gene="bla"
                     /label=AmpR promoter