LOCUS Exported 3689 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pPV576, complete sequence.
ACCESSION KU666001
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3689)
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 3689)
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 3689)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..3689
/organism="Cloning vector pPV576"
/mol_type="other DNA"
/db_xref="taxon:1821234"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 239..257
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
misc_feature join(337..359,484..506)
/label=cherry target sequence
/note="cherry target sequence"
misc_feature join(360..409,434..483)
/label=arms of Daf16
/note="arms of Daf16"
promoter complement(577..595)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(602..618)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 756..1058
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS 1407..2201
/codon_start=1
/product="kanamycin resistance protein"
/label=kanamycin resistance protein
/protein_id="ANB62415.1"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
regulatory 2402..2411
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2408..2782
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
rep_origin 2920..3508
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LOCUS Exported 3689 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pPV576, complete sequence.
ACCESSION KU666001
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3689)
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 3689)
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 3689)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..3689
/organism="Cloning vector pPV576"
/mol_type="other DNA"
/db_xref="taxon:1821234"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 239..257
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
misc_feature join(337..359,484..506)
/label=cherry target sequence
/note="cherry target sequence"
misc_feature join(360..409,434..483)
/label=arms of Daf16
/note="arms of Daf16"
promoter complement(577..595)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(602..618)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 756..1058
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS 1407..2201
/codon_start=1
/product="kanamycin resistance protein"
/label=kanamycin resistance protein
/protein_id="ANB62415.1"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
regulatory 2402..2411
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2408..2782
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
rep_origin 2920..3508
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"