DH4-27 vector (V010022)

Basic Vector Information

      • Vector Name:
      • DH4-27
      • Antibiotic Resistance:
      • Tetracycline
      • Length:
      • 5088 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Huang DC, Holtz WJ, Maharbiz MM.

DH4-27 vector Vector Map

DH4-275088 bp6001200180024003000360042004800pRM5'UTRlambda repressormf-ssrA tag (LVA)5'UTRTetRssrA tag (LVA)rrnB T1 terminatorT7Te terminatorRep101pSC101 orilambda t0 terminatorNeoR/KanR

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

DH4-27 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_600        5088 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector DH4-27, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5088)
  AUTHORS   Huang DC, Holtz WJ, Maharbiz MM.
  TITLE     A genetic bistable switch utilizing nonlinear protein degradation
  JOURNAL   J Biol Eng 6 (1), 9 (2012)
  PUBMED    22776405
REFERENCE   2  (bases 1 to 5088)
  AUTHORS   Huang DC, Holtz WJ, Maharbiz MM.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JUN-2012) Electrical Engineering and Computer 
            Sciences, University of California, Berkeley, 656 Sutardja Dai Hall,
            Berkeley, CA 94720, USA
REFERENCE   3  (bases 1 to 5088)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5088)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Eng"; date: "2012"; volume: "6"; issue: "1"; pages: "9"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-JUN-2012) Electrical Engineering and Computer Sciences, 
            University of California, Berkeley, 656 Sutardja Dai Hall, Berkeley,
            CA 94720, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: GENtle v. 1.9.4
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..82
                     /label=pRM
                     /note="pRM"
                     /regulatory_class="promoter"
     misc_feature    9..26
                     /label=oR1
                     /note="oR1"
     misc_feature    33..50
                     /label=oR2
                     /note="oR2"
     misc_feature    59..65
                     /label=oR3mut
                     /note="oR3mut"
     5'UTR           89..123
     CDS             124..834
                     /label=lambda repressor
                     /note="phage lambda repressor"
     misc_feature    835..915
                     /gene="cI with mf degradation tag"
                     /label=mf-ssrA tag (LVA)
                     /note="mf-ssrA tag (LVA)"
     5'UTR           928..968
     CDS             969..1589
                     /label=TetR
                     /note="tetracycline repressor TetR"
     CDS             1590..1622
                     /label=ssrA tag (LVA)
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
     terminator      1661..1732
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1748..1775
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2309..3256)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     rep_origin      complement(3304..3526)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4018..4112)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4146..4937)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"

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