LOCUS       Exported                4485 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  TF expression vector pB1H1, complete sequence.
ACCESSION   DQ515894
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4485)
  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 4485)
  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 4485)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4485
                     /organism="TF expression vector pB1H1"
                     /mol_type="other DNA"
                     /label=alpha-fused transcription factor expression vector
                     /note="alpha-fused transcription factor expression vector"
                     /db_xref="taxon:382947"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(848..1393)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     promoter        1842..1872
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1880..1896
                     /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     1904..1920
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1966..2709
                     /note="RNA polymerase alpha subunit coding region; used for
                     expressing fusion proteins with transcription factors"
     CDS             2719..2742
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4045..219)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(4045..4482)
                     /codon_start=1
                     /gene="Cm"
                     /product="chloramphenicol acetyltransferase"
                     /label=Cm
                     /protein_id="ABF48728.1"
                     /translation="MAMKDGELVIWDSVHPCYTVFHEQTETFSSLWSEYHDDFRQFLHI
                     YSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANMDNFFAPVFTMGKYYT
                     QGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(4045..4482)
                     /gene="Cm"
                     /label=Cm