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PBAD1030A vector (V009302#)

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Basic Vector Information

Basic Vector Information
Vector Name PBAD1030A Antibiotic Resistance Ampicillin
Length 3586 bp Type Cloning vector
Source Chakravartty V, Cronan JE.

PBAD1030A vector Vector Map

araC araBAD promoter rrnB T1 terminator rrnB T2 terminator AmpR promoter bla

PBAD1030A vector Sequence

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Download GeneBank File(.gb)

LOCUS       Exported                3586 bp ds-DNA     circular SYN 17-DEC-2018
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3586
                     /organism="Cloning vector PBAD1030A"
                     /mol_type="other DNA"
                     /db_xref="taxon:1670622"
     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"
     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

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 ℃.