pETHISglpX vector (V006460)

Basic Vector Information

Vector Name:
pETHISglpX
Antibiotic Resistance:
Kanamycin
Length:
6238 bp
Type:
Expression vector
Replication origin:
ori
Source/Author:
Woolston BM, King JR, Reiter M, Van Hove B, Stephanopoulos G.

pETHISglpX vector Map

pETHISglpX6238 bp30060090012001500180021002400270030003300360039004200450048005100540057006000ropCAP binding sitelacIlacI promoterT7 promoterlac operatorRBS6xHisFructose-1,6-bisphosphatase 1 class 2T7 terminatorf1 oriKanRoribom

pETHISglpX vector Sequence

LOCUS       V006460                 6238 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006460
VERSION     V006460
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6238)
  AUTHORS   Woolston BM, King JR, Reiter M, Van Hove B, Stephanopoulos G.
  TITLE     Improving formaldehyde consumption drives methanol assimilation in
            engineered E. coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6238)
  AUTHORS   Woolston BM, King JR, Reiter M, Van Hove B, Stephanopoulos G.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-APR-2018) Chemical Engineering, Massachusetts
            Institute of Technology, 25 Ames Street, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 6238)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6238)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (02-APR-2018) Chemical Engineering, Massachusetts Institute of
            Technology, 25 Ames Street, Cambridge, MA 02139, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(53..241)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(1016..1037)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1053..2132)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(2133..2210)
                     /label="lacI promoter"
     promoter        2519..2537
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2538..2562
                     /label="lac operator"
                     /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)."
     RBS             2577..2599
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2609..2626
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             2627..3631
                     /gene="glpX"
                     /label="Fructose-1,6-bisphosphatase 1 class 2"
                     /note="Fructose-1,6-bisphosphatase 1 class 2 from
                     Escherichia coli (strain K12). Accession#: P0A9C9"
     terminator      3701..3748
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     rep_origin      3785..4240
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(4336..5148)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      5270..5858
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(6044..6186)
                     /label="bom"
                     /note="basis of mobility region from pBR322"

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