fMCS1 vector (V009985)

Basic Vector Information

      • Vector Name:
      • fMCS1
      • Antibiotic Resistance:
      • Tetracycline
      • Length:
      • 8879 bp
      • Type:
      • Filamentous phage display vector
      • Source/Author:
      • Smith GP, Fernandez A-M.

fMCS1 vector Vector Map

fMCS18879 bp40080012001600200024002800320036004000440048005200560060006400680072007600800084008800pVpVIIpVIIIfd terminatorpVIpImultiple cloning site; contains unique restriction sites for SacI, XhoI, BglII, PstI, PvuII and HindIIITcRtetR/tetA promotersTetR

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

fMCS1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

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"

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