lacI-T7-gusA-pBAV1k vector (V009873)

Basic Vector Information

      • Vector Name:
      • lacI-T7-gusA-pBAV1k
      • Antibiotic Resistance:
      • Kanamycin
      • Length:
      • 6193 bp
      • Type:
      • Expression vector
      • Source/Author:
      • Murin CD, Segal K, Bryksin A, Matsumura I.

lacI-T7-gusA-pBAV1k vector Vector Map

lacI-T7-gusA-pBAV1k6193 bp30060090012001500180021002400270030003300360039004200450048005100540057006000T7 +1

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 ℃.

lacI-T7-gusA-pBAV1k vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6193 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector lacI-T7-gusA-pBAV1k, complete sequence.
ACCESSION   JF828583
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6193)
  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 6193)
  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 6193)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6193
                     /organism="Expression vector lacI-T7-gusA-pBAV1k"
                     /mol_type="other DNA"
                     /db_xref="taxon:1009849"
     misc_feature    1
                     /label=T7 +1
                     /note="T7 +1"
     protein_bind    3..27
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     regulatory      58..64
                     /regulatory_class="ribosome_binding_site"
     CDS             71..1939
                     /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="AEI53604.1"
                     /translation="MGSSHHHHHHSQDPALRAVETPTREIKKLDGLWAFSLDRENCGID
                     QRWWESALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDA
                     VTHYGKVWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVI
                     TDENGKKKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVA
                     NGDVSVELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPL
                     RVGIRSVAVKGEQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSY
                     RTSHYPYAEEMLDWADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQ
                     QAHLQAIKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCV
                     NVMFCDAHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIIT
                     EYGVDTLAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILR
                     VGGNKKGIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQGLCGR"
     gene            71..1939
                     /gene="gusA"
                     /label=gusA
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    1952..1972
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     regulatory      2017..2094
                     /regulatory_class="terminator"
                     /note="t1"
     terminator      2023..2109
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2130..2828)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication initiator protein"
                     /protein_id="AEI53600.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNSSDVIRNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            complement(2130..2828)
                     /gene="repA"
                     /label=repA
     CDS             complement(2895..3056)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORFC"
                     /protein_id="AEI53601.1"
                     /translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
                     RKESEQKK"
     CDS             complement(3097..3306)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORFB"
                     /protein_id="AEI53602.1"
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIVVFMITK"
     repeat_region   3259..3307
                     /label=IRIII
                     /note="IRIII"
     repeat_region   3391..3413
                     /label=IRII
                     /note="IRII"
     CDS             3440..3451
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   3460..3591
                     /label=IRI
                     /note="IRI"
     regulatory      3644..3757
                     /regulatory_class="terminator"
                     /note="t0"
     terminator      3653..3747
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3763..4557)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
                     EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             complement(3763..4425)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase APH (3')-IIIA"
                     /label=aminoglycoside phosphotransferase APH
                     /note="APH(3)(5)-IIIa"
                     /protein_id="AEI53603.1"
                     /translation="MTDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLL
                     MSEADGVLCSEGYEDEQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLN
                     NDLADVDCENWEEDTPFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFI
                     DLGRSGRADKWYDIAFCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    4763..4782
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     CDS             complement(4971..6053)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /protein_id="AEI53605.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(4971..6053)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(6054..6131)
                     /gene="lacI"
                     /label=lacI promoter
     regulatory      6156..6162
                     /regulatory_class="minus_35_signal"
                     /note="from T7"
     promoter        6177..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6177..6193
                     /regulatory_class="promoter"
                     /note="from T7"
     regulatory      6181..6187
                     /regulatory_class="minus_10_signal"
                     /note="from T7"

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