Lauro FM, Eloe EA, Liverani N, Bertoloni G, Bartlett DH.
pFL122 载体载体图谱
pFL122 载体载体序列
LOCUS Exported 8652 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pFL122, complete sequence.
ACCESSION AY785149
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8652)
AUTHORS Lauro FM, Eloe EA, Liverani N, Bertoloni G, Bartlett DH.
TITLE Conjugal vectors for cloning, expression, and insertional
mutagenesis in gram-negative bacteria
JOURNAL BioTechniques 38 (5), 708 (2005)
PUBMED 15945369
REFERENCE 2 (bases 1 to 8652)
AUTHORS Lauro FM, Eloe EA, Liverani N, Bertoloni G, Bartlett DH.
TITLE Direct Submission
JOURNAL Submitted (15-OCT-2004) Marine Biology Research Division, Scripps
Institution of Oceanography, 8750 Biological Grade, La Jolla, CA
92037, USA
REFERENCE 3 (bases 1 to 8652)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8652
/organism="Cloning vector pFL122"
/mol_type="other DNA"
/db_xref="taxon:299444"
CDS 63..866
/codon_start=1
/product="Sm resistance protein A"
/label=Sm resistance protein A
/protein_id="AAV49010.1"
/translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
CDS 866..1702
/codon_start=1
/product="Sm resistance protein B"
/label=Sm resistance protein B
/protein_id="AAV49011.1"
/translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
rep_origin 2346..2740
/label=RSF1010 oriV
/note="replication origin of the broad-host-range plasmid
RSF1010; requires the RSF1010 RepA/B/C proteins for
replication (Scholz et al., 1989)"
CDS complement(2767..3051)
/codon_start=1
/product="mobilization protein C"
/label=mobilization protein C
/protein_id="AAV49012.1"
/translation="MVKGSNKAADRLAKLEEQRARINAEIQRVRAREQQQERKNETRRK
VLVGAMILAKVNSSEWPEDRLMAAMDAYLERDHDRALFGLPPRQKDEPG"
oriT 3082..3169
/label=RSF1010 oriT
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
CDS 3250..5379
/codon_start=1
/product="mobilization protein A"
/label=mobilization protein A
/protein_id="AAV49013.1"
/translation="MAIYHLTAKTGSRSGGQSARAKADYIQREGKYARDMDEVLHAESG
HMPEFVERPADYWDAADLYERANGRLFKEVEFALPVELTLDQQKALASEFAQHLTGAER
LPYTLAIHAGGGENPHCHLMISERINDGIERPAAQWFKRYNGKTPEKGGAQKTEALKPK
AWLEQTREAWADHANRALERAGHDARIDHRTLEAQGIERLPGVHLGPNVVEMEGRGIRT
DRADVALNIDTANAQIIDLQEYREAIDHERNRQSEEIQRHQRVSGADRTAGPEHGDTGR
RSPAGHEPDPAGQRGAGGGVAESPAPDRGGMGGAGQRVAGGSRRGEQRRAERPERVAGV
ALEAMANRDAGFHDAYGGAADRIVALARPDATDNRGRLDLAALGGPMKNDRTLQAIGRQ
LKAMGCERFDIGVRDATTGQMMNREWSAAEVLQNTPWLKRMNAQGNDVYIRPAEQERHG
LVLVDDLSEFDLDDMKAEGREPALVVETSPKNYQAWVKVADAAGGELRGQIARTLASEY
DADPASADSRHYGRLAGFTNRKDKHTTRAGYQPWVLLRESKGKTATAGPALVQQAGQQI
EQAQRQQEKARRLASLELPERQLSRHRRTALDEYRSEMAGLVKRFGDDLSKCDFIAAQK
LASRGRSAEEIGKAMAEASPALAERKPGHEADYIERTVSKVMGLPSVQLARAELARAPA
PRQRGMDRGGPDFSM"
CDS 3998..4411
/codon_start=1
/product="mobilization protein B"
/label=mobilization protein B
/protein_id="AAV49014.1"
/translation="MNAIDRVKKSRGINELAEQIEPLAQSMATLADEARQVMSQTQQAS
EAQAAEWLKAQRQTGAAWVELAKELREVAAEVSSAAQSARSASRGWHWKLWLTVMLASM
MPTVVLLIASLLLLDLTPLTTEDGSIWLRLVAR"
CDS 4408..5379
/codon_start=1
/product="replication protein B"
/label=replication protein B
/protein_id="AAV49015.1"
/translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
CDS 5440..5652
/codon_start=1
/product="unknown protein E"
/label=unknown protein E
/protein_id="AAV49016.1"
/translation="MEYEKSASGSVYLIKSDKGYWLPGGFGYTSNKAEAGRFSVADMAS
LNLDGCTLSLFREDKPFGPGKFLGD"
CDS 5654..5860
/codon_start=1
/product="repressor protein F"
/label=repressor protein F
/protein_id="AAV49017.1"
/translation="MKDQKDKQTGDLLASPDAVRQARYAERMKAKGMRQRKFWLTDDEY
EALRECLEELRAAQGGGSDPASA"
CDS 5890..6729
/codon_start=1
/product="replication protein A"
/label=replication protein A
/protein_id="AAV49018.1"
/translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
CDS 6716..7567
/codon_start=1
/product="replication protein C"
/label=replication protein C
/protein_id="AAV49019.1"
/translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
AG"
CDS complement(7953..8318)
/codon_start=1
/product="LacZ"
/label=LacZ
/note="alpha peptide"
/protein_id="AAV49020.1"
/translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
TDRPSQQLRSLNGEWKL"
primer_bind 8102..8118
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 8128..8146
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 8155..8262
/label=MCS
/note="pBluescript multiple cloning site"
misc_feature complement(8179..8262)
/label=multiple cloning site
/note="multiple cloning site"
primer_bind 8179..8195
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8229..8245)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(8275..8293)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(8314..8330)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8338..8354
/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)."
promoter complement(8362..8392)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8407..8428
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
Lauro FM, Eloe EA, Liverani N, Bertoloni G, Bartlett DH.
pFL122 载体载体图谱
pFL122 载体载体序列
LOCUS Exported 8652 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pFL122, complete sequence.
ACCESSION AY785149
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8652)
AUTHORS Lauro FM, Eloe EA, Liverani N, Bertoloni G, Bartlett DH.
TITLE Conjugal vectors for cloning, expression, and insertional
mutagenesis in gram-negative bacteria
JOURNAL BioTechniques 38 (5), 708 (2005)
PUBMED 15945369
REFERENCE 2 (bases 1 to 8652)
AUTHORS Lauro FM, Eloe EA, Liverani N, Bertoloni G, Bartlett DH.
TITLE Direct Submission
JOURNAL Submitted (15-OCT-2004) Marine Biology Research Division, Scripps
Institution of Oceanography, 8750 Biological Grade, La Jolla, CA
92037, USA
REFERENCE 3 (bases 1 to 8652)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8652
/organism="Cloning vector pFL122"
/mol_type="other DNA"
/db_xref="taxon:299444"
CDS 63..866
/codon_start=1
/product="Sm resistance protein A"
/label=Sm resistance protein A
/protein_id="AAV49010.1"
/translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
CDS 866..1702
/codon_start=1
/product="Sm resistance protein B"
/label=Sm resistance protein B
/protein_id="AAV49011.1"
/translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
rep_origin 2346..2740
/label=RSF1010 oriV
/note="replication origin of the broad-host-range plasmid
RSF1010; requires the RSF1010 RepA/B/C proteins for
replication (Scholz et al., 1989)"
CDS complement(2767..3051)
/codon_start=1
/product="mobilization protein C"
/label=mobilization protein C
/protein_id="AAV49012.1"
/translation="MVKGSNKAADRLAKLEEQRARINAEIQRVRAREQQQERKNETRRK
VLVGAMILAKVNSSEWPEDRLMAAMDAYLERDHDRALFGLPPRQKDEPG"
oriT 3082..3169
/label=RSF1010 oriT
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
CDS 3250..5379
/codon_start=1
/product="mobilization protein A"
/label=mobilization protein A
/protein_id="AAV49013.1"
/translation="MAIYHLTAKTGSRSGGQSARAKADYIQREGKYARDMDEVLHAESG
HMPEFVERPADYWDAADLYERANGRLFKEVEFALPVELTLDQQKALASEFAQHLTGAER
LPYTLAIHAGGGENPHCHLMISERINDGIERPAAQWFKRYNGKTPEKGGAQKTEALKPK
AWLEQTREAWADHANRALERAGHDARIDHRTLEAQGIERLPGVHLGPNVVEMEGRGIRT
DRADVALNIDTANAQIIDLQEYREAIDHERNRQSEEIQRHQRVSGADRTAGPEHGDTGR
RSPAGHEPDPAGQRGAGGGVAESPAPDRGGMGGAGQRVAGGSRRGEQRRAERPERVAGV
ALEAMANRDAGFHDAYGGAADRIVALARPDATDNRGRLDLAALGGPMKNDRTLQAIGRQ
LKAMGCERFDIGVRDATTGQMMNREWSAAEVLQNTPWLKRMNAQGNDVYIRPAEQERHG
LVLVDDLSEFDLDDMKAEGREPALVVETSPKNYQAWVKVADAAGGELRGQIARTLASEY
DADPASADSRHYGRLAGFTNRKDKHTTRAGYQPWVLLRESKGKTATAGPALVQQAGQQI
EQAQRQQEKARRLASLELPERQLSRHRRTALDEYRSEMAGLVKRFGDDLSKCDFIAAQK
LASRGRSAEEIGKAMAEASPALAERKPGHEADYIERTVSKVMGLPSVQLARAELARAPA
PRQRGMDRGGPDFSM"
CDS 3998..4411
/codon_start=1
/product="mobilization protein B"
/label=mobilization protein B
/protein_id="AAV49014.1"
/translation="MNAIDRVKKSRGINELAEQIEPLAQSMATLADEARQVMSQTQQAS
EAQAAEWLKAQRQTGAAWVELAKELREVAAEVSSAAQSARSASRGWHWKLWLTVMLASM
MPTVVLLIASLLLLDLTPLTTEDGSIWLRLVAR"
CDS 4408..5379
/codon_start=1
/product="replication protein B"
/label=replication protein B
/protein_id="AAV49015.1"
/translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
CDS 5440..5652
/codon_start=1
/product="unknown protein E"
/label=unknown protein E
/protein_id="AAV49016.1"
/translation="MEYEKSASGSVYLIKSDKGYWLPGGFGYTSNKAEAGRFSVADMAS
LNLDGCTLSLFREDKPFGPGKFLGD"
CDS 5654..5860
/codon_start=1
/product="repressor protein F"
/label=repressor protein F
/protein_id="AAV49017.1"
/translation="MKDQKDKQTGDLLASPDAVRQARYAERMKAKGMRQRKFWLTDDEY
EALRECLEELRAAQGGGSDPASA"
CDS 5890..6729
/codon_start=1
/product="replication protein A"
/label=replication protein A
/protein_id="AAV49018.1"
/translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
CDS 6716..7567
/codon_start=1
/product="replication protein C"
/label=replication protein C
/protein_id="AAV49019.1"
/translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
AG"
CDS complement(7953..8318)
/codon_start=1
/product="LacZ"
/label=LacZ
/note="alpha peptide"
/protein_id="AAV49020.1"
/translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
TDRPSQQLRSLNGEWKL"
primer_bind 8102..8118
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 8128..8146
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 8155..8262
/label=MCS
/note="pBluescript multiple cloning site"
misc_feature complement(8179..8262)
/label=multiple cloning site
/note="multiple cloning site"
primer_bind 8179..8195
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8229..8245)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(8275..8293)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(8314..8330)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8338..8354
/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)."
promoter complement(8362..8392)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8407..8428
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."