pJG091 vector (V005341)

Basic Vector Information

Vector Name:
pJG091
Antibiotic Resistance:
Ampicillin
Length:
10900 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Goodson J, Klupt S, Zhang C, Straight P, Winkler W.

pJG091 vector Map

pJG09110900 bp5001000150020002500300035004000450050005500600065007000750080008500900095001000010500AmpR promoterAmpRoriSpectinomycin 9-adenylyltransferaseamyE; amyE back; amyE homology for integration into B. amyloliquefaciensChloramphenicol acetyltransferasePdfnA5'UTRlambda tR2 terminatorrrnB T1 terminatormVenusYFP_terminatoramyE-frontLoaPmini-xylAxylO operatorxylO operator-10 (sigA)-35 (sigA)xylRamyE; amyE front; amyE homology for integration into B. amyloliquefaciens

pJG091 vector Sequence

LOCUS       V005341                10900 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005341
VERSION     V005341
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10900)
  AUTHORS   Goodson J, Klupt S, Zhang C, Straight P, Winkler W.
  TITLE     A Broadly Conserved Antiterminator Protein Controls a Regulon of
            Bacillus amyloliquefaciens Antibiotic Gene Clusters
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10900)
  AUTHORS   Goodson JR, Klupt SL.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-JUL-2016) Cell Biology and Molecular Genetics,
            University of Maryland, 3112 Bioscience Research Building, College
            Park, MD 20742, USA
REFERENCE   3  (bases 1 to 10900)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10900)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (01-JUL-2016) Cell Biology and Molecular Genetics, University of
            Maryland, 3112 Bioscience Research Building, College Park, MD 20742,
            USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..107
                     /label="AmpR promoter"
     CDS             108..965
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1139..1727
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(1977..2756)
                     /gene="ant1"
                     /label="Spectinomycin 9-adenylyltransferase"
                     /note="Spectinomycin 9-adenylyltransferase from
                     Staphylococcus aureus (strain N315). Accession#: P0A0D1"
     misc_feature    3865..4865
                     /pseudo=""
                     /note="amyE; amyE back; amyE homology for integration into
                     B. amyloliquefaciens"
     CDS             complement(5239..5886)
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     regulatory      6196..6337
                     /label="PdfnA"
                     /note="PdfnA"
                     /regulatory_class="promoter"
     5'UTR           6345..6745
                     /note="dfnA leader region; dfnA leader region"
     terminator      6745..6792
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     regulatory      6794..6839
                     /label="lambda tR2 terminator"
                     /note="lambda tR2 terminator"
                     /regulatory_class="terminator"
     terminator      complement(6843..6886)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     regulatory      6897..6914
                     /label="Winkler Std. RBS"
                     /note="Winkler Std. RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             6912..7628
                     /label="mVenus"
                     /note="Venus YFP with monomerizing A206K mutation (Nagai et
                     al., 2002; Kremers et al., 2006)"
     regulatory      7638..7671
                     /label="YFP_terminator"
                     /note="YFP_terminator"
                     /regulatory_class="terminator"
     misc_feature    7679..7697
                     /label="amyE-front"
                     /note="amyE-front"
     CDS             complement(7745..8275)
                     /codon_start=1
                     /gene="loaP"
                     /product="LoaP"
                     /label="loaP"
                     /note="LoaP; LoaP NusG-like antiterminator protein"
                     /protein_id="APU90169.1"
                     /translation="MKWYALFVESGKEETVQKFLRLQFDEQALYSIIPKKKVTERKAGI
                     KYEALKKMFPGYVLFKTKMTERTFHKIKELPISCRIVNNGAYYSKERKTYFTTIKDEEI
                     LPIIRLIGEGDTVDYSKVYIENSKVTVASGPLKGMEGIIKKIDKRKRRAKICLSFMGLD
                     KMVNVGIEVLSKP"
     gene            complement(7745..8275)
                     /gene="loaP"
                     /label="loaP"
     CDS             complement(8304..8363)
                     /codon_start=1
                     /gene="mini-xylA"
                     /product="hypothetical protein"
                     /label="mini-xylA"
                     /note="mini xylA ORF"
                     /protein_id="APU90170.1"
                     /translation="MAQSHSSSVNYFVSVNKVV"
     gene            complement(8304..8363)
                     /gene="mini-xylA"
                     /label="mini-xylA"
                     /note="xylA"
     protein_bind    complement(8433..8443)
                     /label="XylR binding site"
                     /bound_moiety="XylR"
                     /note="xylO operator"
     protein_bind    complement(8447..8457)
                     /label="XylR binding site"
                     /bound_moiety="XylR"
                     /note="xylO operator"
     regulatory      complement(8467..8472)
                     /label="-10 (sigA)"
                     /note="-10 (sigA)"
                     /regulatory_class="minus_10_signal"
     regulatory      complement(8490..8497)
                     /label="-35 (sigA)"
                     /note="-35 (sigA)"
                     /regulatory_class="minus_35_signal"
     CDS             8724..9776
                     /codon_start=1
                     /gene="xylR"
                     /product="XylR Xylose Repressor protein"
                     /label="xylR"
                     /note="XylR"
                     /protein_id="APU90171.1"
                     /translation="MTGLNKSTVSSQVNTLMKENLVFEIGQGQSSGGRRPVMLVFNKKA
                     GYSIGIDVGVDYISGILTDLEGTIILDQHHHLESNSPEITKDILIDMIHHFITRMPQSP
                     YGLIGIGICVPGLIDKNQKIVFTPNSNWRDIDLKSFIQEKFNVPVFIENEANAGAYGEK
                     VFGAAKNHNNIIYASISTGIGIGVIINNHLYRGVSGFSGEMGHMTIDFNGPKCSCGNRG
                     CWELYASEKALLKSLQTKEKKVSYQDIIDLAHLNDIGTLNALQNFGFYLGIGLTNILNT
                     FNPQAIILRNSIIESHPMVLNSIRSEVSSRVYPQLGNSYELLPSSLGKNAPALGMSSIV
                     IEHFLDIVKM"
     gene            8724..9776
                     /gene="xylR"
                     /label="xylR"
     misc_feature    9784..10683
                     /pseudo=""
                     /note="amyE; amyE front; amyE homology for integration into
                     B. amyloliquefaciens"

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