araC-pBAD-gusA-pBAV1k vector (V010112)

Basic Vector Information

      • Vector Name:
      • araC-pBAD-gusA-pBAV1k
      • Antibiotic Resistance:
      • Kanamycin
      • Length:
      • 6039 bp
      • Type:
      • Expression vector
      • Source/Author:
      • Murin CD, Segal K, Bryksin A, Matsumura I.

araC-pBAD-gusA-pBAV1k vector Vector Map

araC-pBAD-gusA-pBAV1k6039 bp30060090012001500180021002400270030003300360039004200450048005100540057006000araBAD promoterGusABioBrick suffixt1RepAORFCORFBIRIIFactor Xa siteIRIt0KanRBioBrick prefixAraC

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 ℃.

araC-pBAD-gusA-pBAV1k vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6039 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector araC-pBAD-gusA-pBAV1k, complete sequence.
ACCESSION   JF828585
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6039)
  AUTHORS   Murin CD, Segal K, Bryksin A, Matsumura I.
  TITLE     Expression Vectors for Acinetobacter baylyi ADP1
  JOURNAL   Appl. Environ. Microbiol. 78 (1), 280-283 (2012)
  PUBMED    22020504
REFERENCE   2  (bases 1 to 6039)
  AUTHORS   Murin CD, Segal K, Bryksin A, Matsumura I.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-APR-2011) Biochemistry, Emory University, 1510 Clifton
            Rd NE, Atlanta, GA 30322, USA
REFERENCE   3  (bases 1 to 6039)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6039
                     /organism="Expression vector araC-pBAD-gusA-pBAV1k"
                     /mol_type="other DNA"
                     /db_xref="taxon:1009847"
     promoter        1..285
                     /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)"
     regulatory      4..276
                     /regulatory_class="promoter"
                     /note="from pBAD"
     regulatory      247..253
                     /regulatory_class="minus_35_signal"
     regulatory      271..277
                     /regulatory_class="minus_10_signal"
     misc_feature    286
                     /label=pBAD +1
                     /note="pBAD +1"
     regulatory      304..310
                     /regulatory_class="ribosome_binding_site"
     CDS             319..2187
                     /codon_start=1
                     /gene="gusA"
                     /product="GusA"
                     /label=gusA
                     /note="6-His beta-D-glucuronidase"
                     /db_xref="ASAP:ABE-0005410"
                     /db_xref="EcoGene:EG11055"
                     /db_xref="UniProtKB/Swiss-Prot:P05804"
                     /protein_id="AEI53616.1"
                     /translation="MGSSHHHHHHSQDPALRAVETPTREIKKLDGLWAFSLDRENCGID
                     QRWWESALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDA
                     VTHYGKVWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVI
                     TDENGKKKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVA
                     NGDVSVELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPL
                     RVGIRSVAVKGEQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSY
                     RTSHYPYAEEMLDWADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQ
                     QAHLQAIKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCV
                     NVMFCDAHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIIT
                     EYGVDTLAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILR
                     VGGNKKGIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQGLCGR"
     gene            319..2187
                     /gene="gusA"
                     /label=gusA
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2200..2220
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     regulatory      2265..2329
                     /regulatory_class="terminator"
                     /note="t1"
     terminator      2271..2357
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2378..3076)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication initiator protein"
                     /protein_id="AEI53612.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNSSDVIRNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            complement(2378..3076)
                     /gene="repA"
                     /label=repA
     CDS             complement(3143..3304)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORFC"
                     /protein_id="AEI53613.1"
                     /translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
                     RKESEQKK"
     CDS             complement(3345..3554)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORFB"
                     /protein_id="AEI53614.1"
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIVVFMITK"
     repeat_region   3507..3555
                     /label=IRIII
                     /note="IRIII"
     repeat_region   3639..3661
                     /label=IRII
                     /note="IRII"
     CDS             3688..3699
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   3708..3839
                     /label=IRI
                     /note="IRI"
     regulatory      3892..4005
                     /regulatory_class="terminator"
                     /note="t0"
     terminator      3901..3995
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4011..4805)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
                     EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             complement(4011..4673)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase APH (3')-IIIA"
                     /label=aminoglycoside phosphotransferase APH
                     /note="APH(3)(5)-IIIa"
                     /protein_id="AEI53615.1"
                     /translation="MTDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLL
                     MSEADGVLCSEGYEDEQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLN
                     NDLADVDCENWEEDTPFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFI
                     DLGRSGRADKWYDIAFCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    5011..5030
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     CDS             complement(5135..6013)
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /protein_id="AEI53617.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            complement(5135..6013)
                     /gene="araC"
                     /label=araC

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