Promoter 43 is a constitutive promoter that constitutively expresses the P43 protein in B.subtilis. This promoter has been shown to be recognized and active during the exponential and lag phases of growth. It has been hypothesized that the ability to recognize the promoter in exponential and lag phase of growth is due to the recognition of the promoter by both sigma factor 55 (the major sigma factor A) and sigma factor 37 (the lag phase sigma factor B) 1. The P43 promoter has been previously used for constitutive expression of exogenous genes within B.subtilis vectors 2. The context with which we used the promoter P43 is as a Polymerase Per Second (PoPS) generator.
| 载体基本信息 | |||
| 载体名称 | pHP13-P43 | 载体抗性 | Chloramphenicol |
|---|---|---|---|
| 载体长度 | 5100 bp | 载体类型 | B. subtilis Expression vector |
| 拷贝数 | High copy number | 启动子 | P43 |
| 克隆方法 | Enzyme Cut | ||
pHP13-P43 载体载体图谱
pHP13-P43 载体载体序列
LOCUS Exported 5100 bp ds-DNA circular SYN 17-NOV-2021
DEFINITION Cloning vector pHP13, complete sequence.
ACCESSION DQ297764
VERSION .
KEYWORDS pHP13-P43
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5100)
AUTHORS Jakobsen OM, Benichou A, Flickinger MC, Valla S, Ellingsen TE,
Brautaset T.
TITLE Upregulated transcription of plasmid and chromosomal ribulose
monophosphate pathway genes is critical for methanol assimilation
rate and methanol tolerance in the methylotrophic bacterium Bacillus
methanolicus
JOURNAL J. Bacteriol. 188 (8), 3063-3072 (2006)
PUBMED 16585766
REFERENCE 2 (bases 1 to 5100)
AUTHORS Jakobsen OM, Benichou A, Flickinger MC, Valla S, Ellingsen TE,
Brautaset T.
TITLE Direct Submission
JOURNAL Submitted (18-NOV-2005) Department of Biotechnology, Norwegian
University of Science and Technology, Sem Saelands vei 6/8,
Trondheim 7491, Norway
REFERENCE 3 (bases 1 to 5100)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5100
/organism="Cloning vector pHP13"
/mol_type="other DNA"
/note="vector first presented in: Haima, P., S. Bron, and
G. Venema. 1987. The Effect of Restriction on Shotgun
Cloning and Plasmid Stability in Bacillus-Subtilis Marburg.
Molecular & General Genetics 209:335-342. E. coli/B.
subtilis shuttle vector"
/db_xref="taxon:360321"
regulatory complement(356..415)
/regulatory_class="attenuator"
rep_origin 565..1153
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 1441..1462
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1477..1507
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 1515..1531
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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)."
primer_bind 1539..1555
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 1574..1927
/note="P43"
/note="Promoter P43 is a strong promoter of Bacillus
subtilis expression system. It is derived from cytidine
deaminase (cdd) gene in B. subtilis . It is often used to
express different target proteins."
primer_bind complement(1956..1972)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(2181..2831)
/codon_start=1
/product="chloramphenicol acetyltransferase"
/note="chloramphenicol acetyltransferase"
/note="derived from pC194"
/protein_id="ABC40607.1"
/translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
gene 3257..4279
/gene="rep60"
/note="rep60"
CDS 3257..4279
/codon_start=1
/gene="rep60"
/product="replication protein"
/note="rep60"
/note="involved in initiation of plasmid replication;
derived from pTA1060"
/protein_id="ABC40605.1"
/translation="MIYSSENDYSILEDKTATGKKRDWKGKKRRTNLMAEHYEALESKI
GAPYYGKKAEKLISCAEYLSFKRDPETGKLKLYQAHFCKVRLCPMCAWRRSLKIAYHNK
LIVEEANRQYGCGWIFLTLTIRNVKGERLKPQISAMMEGFRKLFQYKKVKTSVLGFFRA
LEITKNHEEDTYHPHFHVLIPVRKNYFGKNYIKQAEWTSLWKKAMKLDYTPIVDIRRVK
GKAKIDAEQIENDVRNAMMEQKAVLEISKYPVKDTDVVRGNKVTEDNLNTVLYLDDALA
ARRLIGYGGILKEIHKELNLGDAEDGDLVKIEEEDDEVANGAFEVMAYWHPGIKNYIIK
"
CDS complement(4662..296)
/codon_start=1
/product="adenine methylase"
/note="adenine methylase"
/note="derived from pE194; mls resistance"
/protein_id="ABC40606.1"
/translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
LELVKRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDILQFKFPKNQSYKIYGNIPY
NISTDIIRKIVFDSIANEIYLIVEYGFAKRLLNTKRSLALLLMAEVDISILSMVPREYF
HPKPKVNSSLIRLSRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
DLNNISFEQFLSLFNSYKLFNK"
