LOCUS       Exported                7192 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pIPKTA30N, complete sequence.
ACCESSION   EF622218
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7192)
  AUTHORS   Douchkov D, Nowara D, Zierold U, Schweizer P.
  TITLE     A high-throughput gene-silencing system for the functional 
            assessment of defense-related genes in barley epidermal cells
  JOURNAL   Mol. Plant Microbe Interact. 18 (8), 755-761 (2005)
  PUBMED    16134887
REFERENCE   2  (bases 1 to 7192)
  AUTHORS   Douchkov D, Schweizer P.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAY-2007) Cytogenetics, Institute of Plant Genetics 
            and Crop Plant Research (IPK), Corrensstrasse 3, Gatersleben 
            D-06466, Germany
REFERENCE   3  (bases 1 to 7192)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7192
                     /organism="Cloning vector pIPKTA30N"
                     /mol_type="other DNA"
                     /note="dsRNAi vector for transient-induced gene silencing 
                     in plant epidermal cells"
                     /db_xref="taxon:444600"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      230..771
                     /regulatory_class="promoter"
                     /note="CaMV 35S"
     promoter        432..777
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_recomb     791..915
                     /label=attL1
                     /note="attL1"
     protein_bind    791..915
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        940..970
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1024..1683
                     /codon_start=1
                     /product="chloramphenicol resistance protein"
                     /label=chloramphenicol resistance protein
                     /note="CmR"
                     /protein_id="ABQ65198.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2025..2330
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /note="negative selection marker"
                     /protein_id="ABQ65199.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            2025..2330
                     /gene="ccdB"
                     /label=ccdB
     misc_recomb     2371..2495
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(2371..2495)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          2508..2838
                     /number=2
                     /note="RGA2"
     protein_bind    2841..2965
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     2842..2965
                     /label=attR2
                     /note="attR2"
     CDS             complement(3006..3311)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /note="negative selection marker"
                     /protein_id="ABQ65201.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            complement(3006..3311)
                     /gene="ccdB"
                     /label=ccdB
     CDS             complement(3653..4312)
                     /codon_start=1
                     /product="chloramphenicol resistance protein"
                     /label=chloramphenicol resistance protein
                     /note="CmR"
                     /protein_id="ABQ65200.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(4366..4396)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_recomb     4421..4546
                     /label=attR1
                     /note="attR1"
     protein_bind    complement(4421..4545)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      4549..4773
                     /regulatory_class="terminator"
                     /note="CaMV 35S"
     primer_bind     complement(4796..4812)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5286..5390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5391..6251
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ABQ65202.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6422..7010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"