LOCUS Exported 2115 bp ds-DNA circular SYN 18-3-2014
DEFINITION Plasmid with a Zeocin(TM) resistance marker and a TEV cleavage site,
for creating a Gateway(R) entry vector by ligation-independent
cloning (LIC).
ACCESSION .
VERSION .
KEYWORDS pMCentr1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2115)
AUTHORS Midwest Center for Structural Genomics
TITLE Direct Submission
COMMENT For LIC, linearize with SspI and treat with T4 DNA polymerase plus
dGTP.
FEATURES Location/Qualifiers
source 1..2115
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
terminator 268..295
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 387..473
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 537..553
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 569..668
/gene="mutant version of attL"
/bound_moiety="LR Clonase(TM)"
/note="attL1"
/note="recombination site for the Gateway(R) LR reaction"
CDS 671..691
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/note="TEV site"
/translation="ENLYFQS"
protein_bind complement(711..810)
/gene="mutant version of attL"
/bound_moiety="LR Clonase(TM)"
/note="attL2"
/note="recombination site for the Gateway(R) LR reaction"
promoter complement(828..846)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(851..867)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(935..1309)
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/note="BleoR"
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
promoter complement(1328..1375)
/note="EM7 promoter"
/note="synthetic bacterial promoter "
rep_origin 1465..2053
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LOCUS Exported 2115 bp ds-DNA circular SYN 18-3-2014
DEFINITION Plasmid with a Zeocin(TM) resistance marker and a TEV cleavage site,
for creating a Gateway(R) entry vector by ligation-independent
cloning (LIC).
ACCESSION .
VERSION .
KEYWORDS pMCentr1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2115)
AUTHORS Midwest Center for Structural Genomics
TITLE Direct Submission
COMMENT For LIC, linearize with SspI and treat with T4 DNA polymerase plus
dGTP.
FEATURES Location/Qualifiers
source 1..2115
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
terminator 268..295
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 387..473
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 537..553
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 569..668
/gene="mutant version of attL"
/bound_moiety="LR Clonase(TM)"
/note="attL1"
/note="recombination site for the Gateway(R) LR reaction"
CDS 671..691
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/note="TEV site"
/translation="ENLYFQS"
protein_bind complement(711..810)
/gene="mutant version of attL"
/bound_moiety="LR Clonase(TM)"
/note="attL2"
/note="recombination site for the Gateway(R) LR reaction"
promoter complement(828..846)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(851..867)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(935..1309)
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/note="BleoR"
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
promoter complement(1328..1375)
/note="EM7 promoter"
/note="synthetic bacterial promoter "
rep_origin 1465..2053
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"