LOCUS       Exported               10824 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pG2RNAi2, complete sequence.
ACCESSION   KT954097
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10824)
  AUTHORS   Noon JB, Maier TR, Baum TJ.
  TITLE     HgGLAND18 is a malaria-like effector of the soybean cyst nematode 
            that is important for virulence on soybean and efficiently targets 
            host cell nucleoli for suppression of innate immunity
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10824)
  AUTHORS   Noon JB, Maier TR, Baum TJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-OCT-2015) Plant Pathology and Microbiology, Iowa State
            University, 351 Bessey Hall, Ames, IA 50011, USA
REFERENCE   3  (bases 1 to 10824)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10824
                     /organism="Cloning vector pG2RNAi2"
                     /mol_type="other DNA"
                     /db_xref="taxon:1842705"
     misc_feature    114..138
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    complement(115..138)
                     /label=T-DNA right border
                     /note="T-DNA right border"
     primer_bind     334..350
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      432..1348
                     /regulatory_class="promoter"
                     /note="Glycine max polyubiquitin (PGmUBI)"
     misc_feature    1359..1366
                     /label=AscI restriction site
                     /note="AscI restriction site"
     misc_feature    1367..1374
                     /label=SwaI restriction site
                     /note="SwaI restriction site"
     misc_feature    1378..2306
                     /label=GUS linker
                     /note="GUS linker"
     misc_feature    2306..2311
                     /label=BamHI restriction site
                     /note="BamHI restriction site"
     misc_feature    2314..2318
                     /label=AvrII restriction site
                     /note="AvrII restriction site"
     regulatory      2319..2573
                     /regulatory_class="terminator"
                     /note="rbcS"
     regulatory      2917..3434
                     /regulatory_class="promoter"
                     /note="CsVMV"
     CDS             3440..4159
                     /codon_start=1
                     /product="sGFP"
                     /label=sGFP
                     /protein_id="ANC33514.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     regulatory      4174..4453
                     /regulatory_class="terminator"
                     /note="nos"
     terminator      4178..4430
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4472..4488)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4496..4512
                     /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(4520..4550)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4565..4586
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    complement(4762..4787)
                     /label=T-DNA left border
                     /note="T-DNA left border"
     misc_feature    4763..4787
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    5025..5914
                     /label=kanR
                     /note="kanR"
     CDS             5080..5895
                     /codon_start=1
                     /gene="aph(3)-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      5990..6578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      6194..6474
                     /label=pBR322 ori
                     /note="pBR322 ori"
     misc_feature    6614..6874
                     /label=pBR322 bom
                     /note="pBR322 bom"
     misc_feature    6764..6904
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      7248..7442
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    7284..8284
                     /label=pVS1 Rep
                     /note="pVS1 Rep"
     CDS             complement(7508..8575)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     misc_feature    8877..9877
                     /label=pVS1 Sta
                     /note="pVS1 Sta"
     CDS             complement(9009..9638)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"