LOCUS       Exported                4157 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Plant expression vector pGIGI, complete sequence.
ACCESSION   AY720440
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4157)
  AUTHORS   Ferrando A, Koncz-Kalman Z, Farras R, Tiburcio A, Schell J, Koncz C.
  TITLE     Detection of in vivo protein interactions between Snf1-related 
            kinase subunits with intron-tagged epitope-labelling in plants cells
  JOURNAL   Nucleic Acids Res. 29 (17), 3685-3693 (2001)
  PUBMED    11522840
REFERENCE   2  (bases 1 to 4157)
  AUTHORS   Berendzen KW, Koncz C.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-AUG-2004) Development, Max-Planck Institute for Plant 
            Breeding Research, Carl-von-Linne-Weg 10, Cologne D-50829, Germany
REFERENCE   3  (bases 1 to 4157)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4157
                     /organism="Plant expression vector pGIGI"
                     /mol_type="other DNA"
                     /db_xref="taxon:292605"
     misc_feature    1..18
                     /note="Tth111I, NotI, XhoI, KpnI; MCS promoter"
     regulatory      19..758
                     /regulatory_class="promoter"
                     /note="duplicated 35S CaMV (Cabb B-JI isolate); 35SS; 
                     overexpression in plants"
     promoter        409..754
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    759..793
                     /note="HindIII, NcoI, SmaI, XbaI, BglII; MCS upstream"
     misc_feature    794..808
                     /label=N-term c-myc epitope
                     /note="N-term c-myc epitope"
     intron          809..997
                     /note="PIV2 optimized; prevents detection of epitope tag in
                     bacteria"
     misc_feature    998..1012
                     /label=C-term c-myc epitope
                     /note="C-term c-myc epitope"
     misc_feature    1022..1041
                     /note="SalI, BamHI, PstI, EcoRI; MCS downstream"
     regulatory      1042..1294
                     /regulatory_class="terminator"
                     /note="polyadenylation signal from nopaline synthase gene 
                     of the Agrobacterium Ti plasmid; NOS terminator"
     misc_feature    1295..1364
                     /note="KpnI, SacI, NotI, SacI, T7, SfiI; MCS terminator"
     promoter        complement(1326..1344)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(1365..1593)
                     /label=LacI
                     /note="LacI"
     primer_bind     complement(1374..1390)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1398..1414
                     /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(1422..1452)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1467..1488
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1776..2364)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2535..3395)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /note="resistance to ampcillin"
                     /protein_id="AAU21239.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2535..3395)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(3396..3500)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3527..3982
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     4123..4139
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"