LOCUS       Exported                6149 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pNIA-Cb, complete sequence.
ACCESSION   EF587312
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6149)
  AUTHORS   Zaltsman A, Yi BY, Krichevsky A, Gafni Y, Citovsky V.
  TITLE     Yeast-plant coupled vector system for identification of nuclear 
            proteins
  JOURNAL   Plant Physiol. 145 (4), 1264-1271 (2007)
  PUBMED    17704231
REFERENCE   2  (bases 1 to 6149)
  AUTHORS   Zaltsman AA, Citovsky V.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-APR-2007) Biochemistry and Cell Bio, State University 
            of New York, Stony Brook, NY 11777, USA
REFERENCE   3  (bases 1 to 6149)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6149
                     /organism="Yeast expression vector pNIA-Cb"
                     /mol_type="other DNA"
                     /db_xref="taxon:443237"
     CDS             25..1074
                     /codon_start=1
                     /product="LexA-Gal4 chimeric protein"
                     /label=LexA-Gal4 chimeric protein
                     /note="for NLS identification"
                     /protein_id="ABQ43168.1"
                     /translation="MKALTARQQEVFDLIHDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKGLEKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWLEFNFNQSGNIADSSLSFTFT
                     NSSNGPNLITTQTNSQALSQPIASSNVHDNFMNNEITASKIDDGNNSKPLSPGWTDQTA
                     YNAFGITTGMFNTTTMDDVYNYLFDDEDTPPNPKKEIEYPVYKVRTQISSSSFEFCSRR
                     YRGPGIHRI"
     misc_feature    25..630
                     /label=mLexA
                     /note="mLexA"
     CDS             25..630
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5-CTGTATATATATACAG-3."
                     /translation="MKALTARQQEVFDLIHDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKGLEKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     misc_feature    631..984
                     /label=Gal4AD
                     /note="Gal4AD"
     CDS             637..975
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="NFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSNV
                     HDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDDE
                     DTPPNPKKE"
     misc_feature    984..1071
                     /label=MCS
                     /note="MCS"
     misc_feature    998..1063
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1086..1310
                     /regulatory_class="terminator"
                     /note="SV40"
     polyA_signal    1187..1308
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1379..2083)
                     /codon_start=1
                     /product="TRP1"
                     /label=TRP1
                     /protein_id="ABQ43169.1"
                     /translation="MIKHTKAAWSMSVINFTGSSGPLVKVCGLQSTEAAECALDSDADL
                     LGIICVPNRKRTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGI
                     DIVQLHGDESWQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGE
                     LLDWNSISDWVGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDS
                     NKIANFVKNAKK"
     CDS             complement(1379..2053)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(2054..2155)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2169..2185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2193..2209
                     /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(2217..2247)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2262..2283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2571..3159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3330..4190)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /protein_id="ABQ43170.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEAS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3330..4190)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(4191..4295)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4577..5741
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     regulatory      5746..6149
                     /regulatory_class="promoter"
                     /note="ADH1"
     promoter        5751..6147
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"