LOCUS Exported 8879 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Filamentous phage display vector fMCS1, complete sequence.
ACCESSION AF218733
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8879)
AUTHORS Smith GP, Fernandez A-M.
TITLE Direct Submission
JOURNAL Submitted (22-DEC-1999) Division of Biological Sciences, University
of Missouri, Tucker Hall, Columbia, MO 65211-7400, USA
REFERENCE 2 (bases 1 to 8879)
AUTHORS .
TITLE Direct Submission
COMMENT fMCS1 is a filamentous phage cloning vector derived from fd-tet by
replacing 329 non-essential base-pairs with a 25-bp multiple cloning
site. fMCS1 is similar in all important respects to its fd-tet
parent. Minus strand synthesis is slow, so double-stranded RF copy
number is low (about 1 in growing cells; 100 in stationary-phase
cells). Yield of physical particles is about 5e11 virions/ml.
Infection with fMCS1 transduces tetracycline resistance into the
host cell, and the vector can be propagated like a
plasmid by growing infected cells in medium containing tetracycline.
Plaques are very small. Foreign inserts can be spliced into the
unique restriction sites in the multiple cloning site, and are
generally stable.
FEATURES Location/Qualifiers
source join(1..5646,5672..8879)
/organism="Filamentous phage cloning vector fd-tet"
/mol_type="genomic DNA"
/label=presented in GenBank Acession Number AF217317
/note="presented in GenBank Acession Number AF217317"
/db_xref="taxon:127516"
source 1..8879
/organism="Filamentous phage display vector fMCS1"
/focus
/lab_host="Escherichia coli"
/mol_type="genomic DNA"
/db_xref="taxon:127589"
CDS 843..1106
/codon_start=1
/gene="V"
/product="pV"
/label=M13 gene V
/note="replication"
/translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQLCYVDLGNEYPVLV
KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
CDS 1108..1209
/codon_start=1
/gene="VII"
/product="pVII"
/label=M13 gene VII
/note="minor coat protein"
/translation="MEQVADFDTIYQAMIQISVVLCFALGIIAGGQR"
CDS 1206..1304
/codon_start=1
/gene="IX"
/product="pIX"
/label=M13 gene IX
/note="minor coat protein"
/translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
CDS 1301..1522
/codon_start=1
/gene="VIII"
/product="pVIII"
/label=M13 gene VIII
/note="major coat protein"
/translation="MKKSLVLKASVAVATLVPMLSFAAEGDDPAKAAFDSLQASATEYI
GYAWAMVVVIVGATIGIKLFKKFTSKAS"
terminator 1526..1574
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
CDS 2856..3194
/codon_start=1
/gene="VI"
/product="pVI"
/label=M13 gene VI
/note="minor coat protein"
/translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKAAIFIFDVKQKI
VSYLDWDK"
CDS 3197..4243
/codon_start=1
/gene="I"
/product="pI"
/label=M13 gene I
/note="phage assembly"
/translation="MAVYFVTGKLGSGKTLVSVGKIQDKIVAGCKIATNLDLRLQNLPQ
VGRFAKTPRVLRIPDKPSISDLLAIGRGNDSYDENKNGLLVLDECGTWFNTRSWNDKER
QPIIDWFLHARKLGWDIIFLVQDLSIVDKQARSALAEHVVYCRRLDRITLPFVGTLYSL
VTGSKMPLPKLHVGVVKYGDSQLSPTVERWLYTGKNLYNAYDTKQAFSSNYDSGVYSYL
TPYLSHGRYFKPLNLGQKMKLTKIYLKKFSRVLCLAIGFASAFTYSYITQPKPEVKKVV
SQTYDFDKFTIDSSQRLNLSYRYVFKDSKGKLINSDDLQKQGYSITYIDLCTVSIKKGN
SNEIVKCN"
CDS 4221..5501
/codon_start=1
/product="protein IV"
/label=protein IV
/protein_id="AAF78531.1"
/translation="MKLLNVINFVFLMFVSSSSFAQVIEMNNSPLRDFVTWYSKQTGES
VIVSPDVKGTVTVYSSDVKPENLRNFFISVLRANNFDMVGSIPSIIQKYNPNSQDYIDE
LPSSDIQEYDDNSAPSGGFFVPQNDNVTQTFKINNVRAKDLIRVVELFVKSNTSKSSNV
LSVDGSNLLVVSAPKDILDNLPQFLSTVDLPTDQILIEGLIFEVQQGDALDFSFAAGSQ
RGTVAGGVNTDRLTSVLSSAGGSFGIFNGDVLGLSVRALKTNSHSKILSVPRILTLSGQ
KGSISVGQNVPFITGRVTGESANVNNPFQTVERQNVGISMSVFPVAMAGGNIVLDITSK
ADSLSSSTQASDVITNQRSIATTVNLRDGQTLLLGGLTDYKNTSQDSGVPFLSKIPLIG
LLFSSRSDSNEESTLYVLVKATIVRAL"
misc_feature 5646..5671
/note="multiple cloning site; contains unique restriction
sites for SacI, XhoI, BglII, PstI, PvuII and HindIII"
CDS complement(5935..7140)
/codon_start=1
/gene="tetA from transposon Tn10"
/product="tetracycline efflux protein"
/label=TcR
/note="confers resistance to tetracycline"
/translation="MNSSTKIALVITLLDAMGIGLIMPVLPTLLREFIASEDIANHFGV
LLALYALMQVIFAPWLGKMSDRFGRRPVLLLSLIGASLDYLLLAFSSALWMLYLGRLLS
GITGATGAVAASVIADTTSASQRVKWFGWLGASFGLGLIAGPIIGGFAGEISPHSPFFI
AALLNIVTFLVVMFWFRETKNTRDNTDTEVGVETQSNSVYITLFKTMPILLIIYFSAQL
IGQIPATVWVLFTENRFGWNSMMVGFSLAGLGLLHSVFQAFVAGRIATKWGEKTAVLLG
FIADSSAFAFLAFISEGWLVFPVLILLAGGGIALPALQGVMSIQTKSHQQGALQGLLVS
LTNATGVIGPLLFAVIYNHSLPIWDGWIWIIGLAFYCIIILLSMTFMLTPQAQGSKQET
SA"
promoter 7148..7203
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 7149..7167
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 7179..7197
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
CDS 7222..7845
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
CDS 8478..831
/codon_start=1
/gene="II"
/product="pII"
/label=M13 gene II
/note="replication"
/translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGMNLSAR
TVEYHIDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
FGTTDLALCSEALLLNFANSLPCLYDLLDVNATTISRIDATFSARAPNENIAKQVIDHL
RNVSNGQTKSTRSQNWESTVTWNETSRHRTLVAYLKHVELQHQIQQLSSKPSAKMTSYQ
KEQLKVLSNPDLLEFASGLVRFEARIETRYLKSFGLPLNLFDAIRFASDYNRQGKDLIF
DLWSFSFSELFKAFEGDSMNIYDDSAVLDAIQSKHFTITPSGKTSFAKASRYFGFYRRL
VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
DWYNEPVLKIA"
LOCUS Exported 8879 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Filamentous phage display vector fMCS1, complete sequence.
ACCESSION AF218733
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8879)
AUTHORS Smith GP, Fernandez A-M.
TITLE Direct Submission
JOURNAL Submitted (22-DEC-1999) Division of Biological Sciences, University
of Missouri, Tucker Hall, Columbia, MO 65211-7400, USA
REFERENCE 2 (bases 1 to 8879)
AUTHORS .
TITLE Direct Submission
COMMENT fMCS1 is a filamentous phage cloning vector derived from fd-tet by
replacing 329 non-essential base-pairs with a 25-bp multiple cloning
site. fMCS1 is similar in all important respects to its fd-tet
parent. Minus strand synthesis is slow, so double-stranded RF copy
number is low (about 1 in growing cells; 100 in stationary-phase
cells). Yield of physical particles is about 5e11 virions/ml.
Infection with fMCS1 transduces tetracycline resistance into the
host cell, and the vector can be propagated like a
plasmid by growing infected cells in medium containing tetracycline.
Plaques are very small. Foreign inserts can be spliced into the
unique restriction sites in the multiple cloning site, and are
generally stable.
FEATURES Location/Qualifiers
source join(1..5646,5672..8879)
/organism="Filamentous phage cloning vector fd-tet"
/mol_type="genomic DNA"
/label=presented in GenBank Acession Number AF217317
/note="presented in GenBank Acession Number AF217317"
/db_xref="taxon:127516"
source 1..8879
/organism="Filamentous phage display vector fMCS1"
/focus
/lab_host="Escherichia coli"
/mol_type="genomic DNA"
/db_xref="taxon:127589"
CDS 843..1106
/codon_start=1
/gene="V"
/product="pV"
/label=M13 gene V
/note="replication"
/translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQLCYVDLGNEYPVLV
KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
CDS 1108..1209
/codon_start=1
/gene="VII"
/product="pVII"
/label=M13 gene VII
/note="minor coat protein"
/translation="MEQVADFDTIYQAMIQISVVLCFALGIIAGGQR"
CDS 1206..1304
/codon_start=1
/gene="IX"
/product="pIX"
/label=M13 gene IX
/note="minor coat protein"
/translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
CDS 1301..1522
/codon_start=1
/gene="VIII"
/product="pVIII"
/label=M13 gene VIII
/note="major coat protein"
/translation="MKKSLVLKASVAVATLVPMLSFAAEGDDPAKAAFDSLQASATEYI
GYAWAMVVVIVGATIGIKLFKKFTSKAS"
terminator 1526..1574
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
CDS 2856..3194
/codon_start=1
/gene="VI"
/product="pVI"
/label=M13 gene VI
/note="minor coat protein"
/translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKAAIFIFDVKQKI
VSYLDWDK"
CDS 3197..4243
/codon_start=1
/gene="I"
/product="pI"
/label=M13 gene I
/note="phage assembly"
/translation="MAVYFVTGKLGSGKTLVSVGKIQDKIVAGCKIATNLDLRLQNLPQ
VGRFAKTPRVLRIPDKPSISDLLAIGRGNDSYDENKNGLLVLDECGTWFNTRSWNDKER
QPIIDWFLHARKLGWDIIFLVQDLSIVDKQARSALAEHVVYCRRLDRITLPFVGTLYSL
VTGSKMPLPKLHVGVVKYGDSQLSPTVERWLYTGKNLYNAYDTKQAFSSNYDSGVYSYL
TPYLSHGRYFKPLNLGQKMKLTKIYLKKFSRVLCLAIGFASAFTYSYITQPKPEVKKVV
SQTYDFDKFTIDSSQRLNLSYRYVFKDSKGKLINSDDLQKQGYSITYIDLCTVSIKKGN
SNEIVKCN"
CDS 4221..5501
/codon_start=1
/product="protein IV"
/label=protein IV
/protein_id="AAF78531.1"
/translation="MKLLNVINFVFLMFVSSSSFAQVIEMNNSPLRDFVTWYSKQTGES
VIVSPDVKGTVTVYSSDVKPENLRNFFISVLRANNFDMVGSIPSIIQKYNPNSQDYIDE
LPSSDIQEYDDNSAPSGGFFVPQNDNVTQTFKINNVRAKDLIRVVELFVKSNTSKSSNV
LSVDGSNLLVVSAPKDILDNLPQFLSTVDLPTDQILIEGLIFEVQQGDALDFSFAAGSQ
RGTVAGGVNTDRLTSVLSSAGGSFGIFNGDVLGLSVRALKTNSHSKILSVPRILTLSGQ
KGSISVGQNVPFITGRVTGESANVNNPFQTVERQNVGISMSVFPVAMAGGNIVLDITSK
ADSLSSSTQASDVITNQRSIATTVNLRDGQTLLLGGLTDYKNTSQDSGVPFLSKIPLIG
LLFSSRSDSNEESTLYVLVKATIVRAL"
misc_feature 5646..5671
/note="multiple cloning site; contains unique restriction
sites for SacI, XhoI, BglII, PstI, PvuII and HindIII"
CDS complement(5935..7140)
/codon_start=1
/gene="tetA from transposon Tn10"
/product="tetracycline efflux protein"
/label=TcR
/note="confers resistance to tetracycline"
/translation="MNSSTKIALVITLLDAMGIGLIMPVLPTLLREFIASEDIANHFGV
LLALYALMQVIFAPWLGKMSDRFGRRPVLLLSLIGASLDYLLLAFSSALWMLYLGRLLS
GITGATGAVAASVIADTTSASQRVKWFGWLGASFGLGLIAGPIIGGFAGEISPHSPFFI
AALLNIVTFLVVMFWFRETKNTRDNTDTEVGVETQSNSVYITLFKTMPILLIIYFSAQL
IGQIPATVWVLFTENRFGWNSMMVGFSLAGLGLLHSVFQAFVAGRIATKWGEKTAVLLG
FIADSSAFAFLAFISEGWLVFPVLILLAGGGIALPALQGVMSIQTKSHQQGALQGLLVS
LTNATGVIGPLLFAVIYNHSLPIWDGWIWIIGLAFYCIIILLSMTFMLTPQAQGSKQET
SA"
promoter 7148..7203
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 7149..7167
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 7179..7197
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
CDS 7222..7845
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
CDS 8478..831
/codon_start=1
/gene="II"
/product="pII"
/label=M13 gene II
/note="replication"
/translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGMNLSAR
TVEYHIDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
FGTTDLALCSEALLLNFANSLPCLYDLLDVNATTISRIDATFSARAPNENIAKQVIDHL
RNVSNGQTKSTRSQNWESTVTWNETSRHRTLVAYLKHVELQHQIQQLSSKPSAKMTSYQ
KEQLKVLSNPDLLEFASGLVRFEARIETRYLKSFGLPLNLFDAIRFASDYNRQGKDLIF
DLWSFSFSELFKAFEGDSMNIYDDSAVLDAIQSKHFTITPSGKTSFAKASRYFGFYRRL
VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
DWYNEPVLKIA"