LOCUS       40924_7441        3827 bp DNA     circular SYN 17-DEC-2018
DEFINITION  RNA interference vector pBSK-Gus, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3827)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "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"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     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"
     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"
     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"
     protein_bind    complement(1702..1718)
                     /label=lac operator
                     /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    complement(1771..1792)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2080..2668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2842..3699)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3700..3804)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattccgcc gggaatggtg
      721 attaccgacg aaaacggcaa gaaaaagcag tcttacttcc atgatttctt taactatgcc
      781 ggaatccatc gcagcgtaat gctctacacc acgccgaaca cctgggtgga cgatatcacc
      841 gtggtgacgc atgtcgcgca agactgtaac cacgcgtctg ttgactggca ggtggtggcc
      901 aatggtgatg tcagcgttga actgcgtgat gcggatcaac aggtggttgc aactggacaa
      961 ggcactagcg ggactttgca agtggtgaat ccgcacctct ggcaaccggg tgaaggttat
     1021 ctctatgaac tgtgcgtcac agccaaaagc cagacagagt gtgatatcta cccgcttcgc
     1081 gtcggcatcc ggtcagtggc agtgaagggc gaacagttcc tgattaacca caaaccgttc
     1141 tactttactg gctttggtcg tcatgaagat gcggacttgc gtggcaaagg attcgataac
     1201 gtgctgatgg tgcacgacca cgcattaatg gactggattg gggccaactc ctaccgtacc
     1261 tcgcattacc cttacgctga agagatgctc gactgggcag atgaacatgg catcgtggtg
     1321 attgacgaaa ctgctgctgt cggctttaac ctctctttag gcattggttt cgaagcgggc
     1381 aacaagccga aagaactgta cagcgaagag gcagtcaacg gggaaactca gcaagcgcac
     1441 ttacaggcga ttaaagagct gatagcgcgt gacaaaaacc acccaagcgt ggtgatgtgg
     1501 agtattgcca acgaaccgga tacccgtccg caaggtgcac gggaatattt cgcgccactg
     1561 gcggaagcaa cgcgtaaact cgacggatcc actagttcta gagcggccgc caccgcggtg
     1621 gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     1681 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1741 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1801 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1861 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1921 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2521 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2641 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2761 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2821 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2881 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2941 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3001 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3061 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3121 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3181 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3241 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3301 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3361 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3421 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3481 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3541 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3601 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3661 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3721 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3781 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
