LOCUS       Exported                4980 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  TF expression vector pB1H2, complete sequence.
ACCESSION   DQ515895
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4980)
  AUTHORS   Meng X, Brodsky MH, Wolfe SA.
  TITLE     A bacterial one-hybrid system for determining the DNA-binding 
            specificity of transcription factors
  JOURNAL   Nat. Biotechnol. 23 (8), 988-994 (2005)
  PUBMED    16041365
REFERENCE   2  (bases 1 to 4980)
  AUTHORS   Meng X, Brodsky MH, Wolfe SA.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-APR-2006) Program in Gene Function and Expression, 
            University of Massachusetts Medical School, 364 Plantation St., 
            Worcester, MA 01605, USA
REFERENCE   3  (bases 1 to 4980)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4980
                     /organism="TF expression vector pB1H2"
                     /mol_type="other DNA"
                     /label=alpha-fused transcription factor expression vector
                     /note="alpha-fused transcription factor expression vector"
                     /db_xref="taxon:382948"
     promoter        89..119
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    127..143
                     /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     151..167
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    213..956
                     /note="RNA polymerase alpha subunit coding region; used for
                     expressing fusion proteins with transcription factors"
     CDS             966..989
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      2041..2496
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2522..2626
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2627..3487
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ABF48729.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2627..3487
                     /gene="bla"
                     /label=bla
     rep_origin      3658..4246
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4432..4572
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4674..4865)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"