LOCUS Exported 7535 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pPV505, complete sequence.
ACCESSION KU665996
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7535)
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 7535)
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 7535)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7535
/organism="Cloning vector pPV505"
/mol_type="other DNA"
/db_xref="taxon:1821229"
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="ANB62407.1"
/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 2949..4150
/regulatory_class="promoter"
/note="Act2; promoter"
CDS complement(3315..3326)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
misc_feature 4259..5124
/label=similar to green fluorescent protein
/note="similar to green fluorescent protein"
misc_feature 5125..5227
/note="P2A; linker"
CDS 5159..5215
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 5228..5926
/codon_start=1
/product="red fluorescent protein"
/label=red fluorescent protein
/note="RFP"
/protein_id="ANB62408.1"
/translation="MASSEDVIKEFMRFKVKMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERIMNFEDGGV
VTVTQDSTLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYKAKKQVQLPGAYKTDIKLDIISHNEDYTIVEQYERAEGRHS
TGAGSPRNLF"
misc_feature 5939..6501
/note="Era1; UTR"
primer_bind 6515..6531
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6920..6936)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 7078..7533
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
LOCUS Exported 7535 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pPV505, complete sequence.
ACCESSION KU665996
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7535)
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 7535)
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 7535)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7535
/organism="Cloning vector pPV505"
/mol_type="other DNA"
/db_xref="taxon:1821229"
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="ANB62407.1"
/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 2949..4150
/regulatory_class="promoter"
/note="Act2; promoter"
CDS complement(3315..3326)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
misc_feature 4259..5124
/label=similar to green fluorescent protein
/note="similar to green fluorescent protein"
misc_feature 5125..5227
/note="P2A; linker"
CDS 5159..5215
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 5228..5926
/codon_start=1
/product="red fluorescent protein"
/label=red fluorescent protein
/note="RFP"
/protein_id="ANB62408.1"
/translation="MASSEDVIKEFMRFKVKMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERIMNFEDGGV
VTVTQDSTLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYKAKKQVQLPGAYKTDIKLDIISHNEDYTIVEQYERAEGRHS
TGAGSPRNLF"
misc_feature 5939..6501
/note="Era1; UTR"
primer_bind 6515..6531
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6920..6936)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 7078..7533
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"