LOCUS       Exported                3827 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  RNA interference vector pBSK-Gus, complete sequence.
ACCESSION   HQ343202
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3827)
  AUTHORS   Narwade AV, Chakrabarty PK, Kalbande BB, Chavhan RL, Warade JG.
  TITLE     Hairpin RNA generating vector designed by cloning 878 bp Gus 
            fragment from E. coli GUSA (N358Q) amplified from pCAMBIA-1302 as 
            stuffer. The vector can be utilized for generating dsRNA of a given 
            sequence by cloning the targeted DNA in sense and antisense 
            orientations flanking the stuffer
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3827)
  AUTHORS   Narwade AV, Chakrabarty PK, Kalbande BB, Chavhan RL, Warade JG.
  TITLE     RNA interference vector pBSK-Gus, to generate inverted repeat of a 
            given sequence
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 3827)
  AUTHORS   Chakrabarty PK, Narwade AV, Kalbande BB, Chavhan RL, Warade JG.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-OCT-2010) Division of Crop Improvement, Central 
            Institute for Cotton Research, Post Bag No. 02, Shankar Nagar P.O., 
            Nagpur, Maharashtra 440010, India
REFERENCE   4  (bases 1 to 3827)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3827
                     /organism="RNA interference vector pBSK-Gus"
                     /mol_type="other DNA"
                     /note="designed to generate inverted repeats (hairpin RNA) 
                     for RNA interference to generate dsRNA by cloning the 
                     targeted DNA in sense and antisense orientations flanking 
                     the stuffer fragment"
                     /db_xref="taxon:933838"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      135..441
                     /label=F1(+) origin of replication
                     /note="F1(+) origin of replication"
     gene            460..706
                     /gene="lacZ-alpha"
                     /label=lacZ-alpha
                     /note="beta-galactosidase alpha fragment; disrupted by 
                     insertion of gusA 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..706
                     /note="MCS; multiple cloning site; derived from pBluescript
                     II SK(+)"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    701..706
                     /label=EcoRI site
                     /note="EcoRI site"
     gene            707..1584
                     /gene="gusA"
                     /label=gusA
                     /note="beta-glucuronidase; stuffer fragment for RNA 
                     interference; derived from Escherichia coli vector 
                     pCAMBIA-1302; M358Q GusA fragment"
     gene            1585..1682
                     /gene="lacZ-alpha"
                     /label=lacZ-alpha
                     /note="beta-galactosidase alpha fragment; disrupted by gusA
                     fragment insertion"
     misc_feature    1585..1626
                     /note="MCS; multiple cloning site; derived from pBluescript
                     II SK(+)"
     misc_feature    1585..1590
                     /label=BamHI site
                     /note="BamHI site"
     primer_bind     complement(1586..1602)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1639..1657)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1678..1694)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1683..1804
                     /regulatory_class="promoter"
                     /note="lac promoter"
     protein_bind    1702..1718
                     /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(1726..1756)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1771..1792
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2024..2691
                     /label=pUC origin of replication
                     /note="pUC origin of replication"
     rep_origin      complement(2080..2668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2839..3699)
                     /codon_start=1
                     /gene="amp"
                     /product="beta-lactamase"
                     /label=amp
                     /protein_id="ADR82071.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2839..3699)
                     /gene="amp"
                     /label=amp
     promoter        complement(3700..3804)
                     /gene="bla"
                     /label=AmpR promoter