LOCUS Exported 4595 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pPV567, complete sequence.
ACCESSION KU665999
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4595)
AUTHORS Lok JB, Shao H, Massey HC, Li X.
TITLE Transgenesis in Strongyloides and related parasitic nematodes:
historical perspectives, current functional genomic applications and
progress towards gene disruption and editing
JOURNAL Parasitology (2016) In press
PUBMED 27000743
REFERENCE 2 (bases 1 to 4595)
AUTHORS Lok JB, Shao H, Massey HC Jr., Li X.
TITLE Direct Submission
JOURNAL Submitted (02-FEB-2016) Pathobiology, University of Pennsylvania,
3800 Spruce Street, Philadelphia, PA 19104, USA
REFERENCE 3 (bases 1 to 4595)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..4595
/organism="Cloning vector pPV567"
/mol_type="other DNA"
/db_xref="taxon:1821232"
promoter 24..128
/gene="bla"
/label=AmpR promoter
CDS 129..989
/codon_start=1
/product="beta-lactamase"
/label=beta-lactamase
/note="Amp-r"
/protein_id="ANB62413.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1160..1748
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2036..2057
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2072..2102
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2110..2126
/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)."
primer_bind 2134..2150
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
regulatory 2956..3467
/regulatory_class="promoter"
/note="U6; Strogyloides ratti promoter"
misc_feature 3468..3490
/note="Daf16; gRNA target"
misc_feature 3491..3573
/label=gRNA scaffold
/note="gRNA scaffold"
misc_RNA 3491..3566
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind 3575..3591
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3980..3996)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 4138..4593
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
LOCUS Exported 4595 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pPV567, complete sequence.
ACCESSION KU665999
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4595)
AUTHORS Lok JB, Shao H, Massey HC, Li X.
TITLE Transgenesis in Strongyloides and related parasitic nematodes:
historical perspectives, current functional genomic applications and
progress towards gene disruption and editing
JOURNAL Parasitology (2016) In press
PUBMED 27000743
REFERENCE 2 (bases 1 to 4595)
AUTHORS Lok JB, Shao H, Massey HC Jr., Li X.
TITLE Direct Submission
JOURNAL Submitted (02-FEB-2016) Pathobiology, University of Pennsylvania,
3800 Spruce Street, Philadelphia, PA 19104, USA
REFERENCE 3 (bases 1 to 4595)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..4595
/organism="Cloning vector pPV567"
/mol_type="other DNA"
/db_xref="taxon:1821232"
promoter 24..128
/gene="bla"
/label=AmpR promoter
CDS 129..989
/codon_start=1
/product="beta-lactamase"
/label=beta-lactamase
/note="Amp-r"
/protein_id="ANB62413.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1160..1748
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2036..2057
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2072..2102
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2110..2126
/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)."
primer_bind 2134..2150
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
regulatory 2956..3467
/regulatory_class="promoter"
/note="U6; Strogyloides ratti promoter"
misc_feature 3468..3490
/note="Daf16; gRNA target"
misc_feature 3491..3573
/label=gRNA scaffold
/note="gRNA scaffold"
misc_RNA 3491..3566
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind 3575..3591
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3980..3996)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 4138..4593
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"