f8-1 vector (V009990)

Basic Vector Information

      • Vector Name:
      • f8-1
      • Antibiotic Resistance:
      • Tetracycline
      • Length:
      • 9183 bp
      • Type:
      • Filamentous phage display vector
      • Source/Author:
      • Smith GP, Fernandez A-M.

f8-1 vector Vector Map

f8-19183 bp40080012001600200024002800320036004000440048005200560060006400680072007600800084008800pVpVIIVIIIfd terminatorpVIpITcRtetR/tetA promotersTetRpII

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 ℃.

f8-1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                9183 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Filamentous phage display vector f8-1, complete sequence.
ACCESSION   AF218734
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9183)
  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 9183)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     Type-8 filamentous phage display vector constructed by engineering 
            three single-base-pair substitutions into Filamentous phage cloning 
            vector fd-tet presented in GenBank Accession Number AF217317. f8-1 
            is very similar in all important characteristics to its fd-tet 
            parent. It is intended as a phage-display  vector. Thus the 9-bp 
            'stuffer' between the PstI and BamHI cloning sites can be replaced 
            with a short degenerate coding sequence to create a library of phage
            displaying different random peptides at the N-terminus of all 3900  
            copies of the major coat protein pVIII.
FEATURES             Location/Qualifiers
     source          1..9183
                     /organism="Filamentous phage display vector f8-1"
                     /lab_host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:127588"
     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="major coat protein pVIII mutant precursor"
                     /label=VIII
                     /protein_id="AAF78532.1"
                     /translation="MKKSLVLKASVAVATLVPMLSFAAEGEDPAKAAFDSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     gene            1301..1522
                     /gene="VIII"
                     /label=VIII
     sig_peptide     1301..1369
                     /gene="VIII"
     misc_feature    1368..1373
                     /label=PstI site
                     /note="PstI site"
     mat_peptide     1370..1519
                     /gene="VIII"
                     /product="major coat protein pVIII mutant"
     variation       1372
                     /replace="t"
                     /label=creates PstI site
                     /note="creates PstI site"
     misc_feature    1381..1386
                     /label=BamHI site
                     /note="BamHI site"
     variation       1381
                     /replace="c"
                     /note="creates BamHI site; results in aspartic acid to 
                     glutamic acid substitution"
     terminator      1526..1574
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     variation       2220
                     /replace="g"
                     /label=removes BamHI site
                     /note="removes BamHI site"
     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             complement(6239..7444)
                     /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        7452..7507
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    7453..7471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7483..7501
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             7526..8149
                     /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             8782..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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