pET15b-vioB vector (V006489)

Basic Vector Information

Vector Name:
pET15b-vioB
Antibiotic Resistance:
Ampicillin
Length:
8684 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Pardee K, Slomovic S, Nguyen PQ, Lee JW, Donghia N, Burrill D, Ferrante T, McSorley F, Furuta Y, Lewandowski M, Boddy CN, Joshi NS, Collins JJ.

pET15b-vioB vector Map

pET15b-vioB8684 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400AmpR promoterAmpRoribomropCAP binding sitelacIlacI promoterT7 promoterlac operatorRBS2-imino-3-(indol-3-yl)propanoate dimeraseFLAGT7 terminatortet promoter

pET15b-vioB vector Sequence

LOCUS       V006489                 8684 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006489
VERSION     V006489
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8684)
  AUTHORS   Pardee K, Slomovic S, Nguyen PQ, Lee JW, Donghia N, Burrill D,
            Ferrante T, McSorley F, Furuta Y, Lewandowski M, Boddy CN, Joshi NS,
            Collins JJ.
  TITLE     Portable, On-demand Biomolecular Manufacturing
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8684)
  AUTHORS   Pardee K, Slomovic S, Nguyen PQ, Lee JW, Donghia N, Burrill D,
            Ferrante T, McSorley F, Furuta Y, Lewandowski M, Boddy CN, Joshi NS,
            Collins JJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUN-2016) Wyss Institute, Harvard University, 3
            Blackfan Circle, Wyss Institute Rm517-1D, Boston, MA 02115, USA
REFERENCE   3  (bases 1 to 8684)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8684)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (29-JUN-2016) Wyss Institute, Harvard University, 3 Blackfan Circle,
            Wyss Institute Rm517-1D, Boston, MA 02115, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        104..208
                     /label="AmpR promoter"
     CDS             209..1066
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1240..1828
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2014..2156)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2261..2449)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3727..3748)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3764..4843)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(4844..4921)
                     /label="lacI promoter"
     promoter        5230..5248
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5249..5273
                     /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             5288..5310
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5317..8310
                     /gene="vioB"
                     /label="2-imino-3-(indol-3-yl)propanoate dimerase"
                     /note="2-imino-3-(indol-3-yl)propanoate dimerase from
                     Chromobacterium violaceum (strain ATCC 12472 / DSM 30191 /
                     JCM 1249 / NBRC 12614 / NCIMB 9131 / NCTC 9757).
                     Accession#: Q9S3V0"
     CDS             8320..8343
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     terminator      8425..8472
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     promoter        complement(8647..8675)
                     /label="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"

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