LOCUS       Exported                5070 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLS3, complete sequence.
ACCESSION   AF169189
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5070)
  AUTHORS   Hays LB, Chen Y-S.A., Hu JC.
  TITLE     A two-hybrid system for characterization of protein-protein 
            interactions in E. coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5070)
  AUTHORS   Hays LB, Chen Y-S.A., Hu JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-JUL-1999) Biochemistry & Biophysics, Texas A&M 
            University, MS2128, College Station, TX 77843-2128, USA
REFERENCE   3  (bases 1 to 5070)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5070
                     /organism="Cloning vector pLS3"
                     /lab_host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /note="bait plasmid for two-hybrid system based on 
                     cooperative binding to adjacent operators; contains pBR322 
                     based origin of replication and ampicillin resistance and 
                     M14 ssDNA origin of replication from pZ150"
                     /db_xref="taxon:101857"
     regulatory      28..89
                     /regulatory_class="promoter"
                     /note="lacUV5 promoter/lac operator"
     promoter        28..58
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    66..82
                     /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)."
     CDS             109..525
                     /codon_start=1
                     /product="434-GCN4-linker"
                     /label=434-GCN4-linker
                     /note="fusion protein; contains N-terminal DNA binding 
                     domain from bacteriophage 434 repressor, a modified leucine
                     zipper from GCN4 and a linker for fusion to additional 
                     protein domains"
                     /protein_id="AAD50896.1"
                     /translation="MSISSRVKSKRIQLGLNQAELAQKVGTTQQSIEQLENGKTKRPRF
                     LPELASALGVSVDWLLNGTSDSNVRFVGHVEPKGKKLRTFTKGDAERWVSTHMKQLEDK
                     IEELLSKNYHLENEIARLKKLIGERGGQLNASGS"
     terminator      599..646
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             2131..2322
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    2424..2564
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2750..3338)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3449..3962
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(4004..4864)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4865..4969)
                     /gene="bla"
                     /label=AmpR promoter