PBAD1030A vector (V009302)

COA reports Sequencing Results MSDS Download GenBank File(.gb)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V009302	PBAD1030A	In stock, instant shipping	Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:: PBAD1030A

Antibiotic Resistance:: Ampicillin

Length:: 3586 bp

Type:: Cloning vector

Replication origin:: RSF ori

Source/Author:: Chakravartty V, Cronan JE.

Promoter:: araBAD

Growth Strain(s):: Top10

Growth Temperature:: 37℃

PBAD1030A vector Map

Copy Sequence View Map

Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

PBAD1030A vector Sequence

LOCUS       Exported                3586 bp DNA     circular SYN 29-NOV-2023
DEFINITION  Cloning vector PBAD1030A, complete sequence.
ACCESSION   KP899255
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3586)
  AUTHORS   Chakravartty V, Cronan JE.
  TITLE     A series of medium and high copy number arabinose-inducible 
            Escherichia coli expression vectors compatible with pBR322 and 
            pACYC184
  JOURNAL   Plasmid 81, 21-26 (2015)
  PUBMED    26021570
REFERENCE   2  (bases 1 to 3586)
  AUTHORS   Chakravartty V, Cronan JE.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAR-2015) Microbiology, University of Illinois, 601S. 
            Goodwin Ave, Urbana, IL 61801, USA
REFERENCE   3  (bases 1 to 3586)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3586)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid 81,
            21-26 (2015)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-MAR-2015) Microbiology, University of Illinois, 601S. Goodwin 
            Ave, Urbana, IL 61801, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3586
                     /mol_type="other DNA"
                     /db_xref="taxon:1670622"
                     /organism="Cloning vector PBAD1030A"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /note="transcription factor"
                     /protein_id="AKM76960.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            complement(96..974)
                     /gene="araC"
                     /label=araC
     promoter        1001..1285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     terminator      1569..1655
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1747..1774
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     misc_feature    complement(1834..2462)
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin 
                     ofreplication can be propagated in E. coli cells that 
                     contain additional plasmids with compatible origins."
     promoter        2479..2583
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2584..3444
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="AKM76961.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2584..3444
                     /gene="bla"
                     /label=bla