f8-5 vector (V009989)

Basic Vector Information

      • Vector Name:
      • f8-5
      • Antibiotic Resistance:
      • Tetracycline
      • Length:
      • 9183 bp
      • Type:
      • Filamentous phage display vector
      • Source/Author:
      • Petrenko VA, Smith GP, Mazooji MM, Quinn T.

f8-5 vector Vector Map

f8-59183 bp40080012001600200024002800320036004000440048005200560060006400680072007600800084008800replication protein pXcytosolic single-stranded DNA binding protein pVminor coat protein pVIImajor coat protein pVIII precursorfd terminatoradsorption protein pIII precursorminor coat protein pVImorphogenetic protein pITn10 tetracycline resistance protein TetAtetR/tetA promotersTn10 tetracycline repressor protein TetR

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 ℃.

f8-5 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                9183 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Filamentous phage display vector f8-5, complete sequence.
ACCESSION   AF464138
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9183)
  AUTHORS   Petrenko VA, Smith GP, Mazooji MM, Quinn T.
  TITLE     An Alpha-Helically Constrained Phage Display Library
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9183)
  AUTHORS   Petrenko VA, Smith GP.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-DEC-2001) Pathobiology, Auburn University, 253 Greene 
            Hall, Auburn, AL 36849, USA
REFERENCE   3  (bases 1 to 9183)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9183
                     /organism="Filamentous phage display vector f8-5"
                     /mol_type="genomic DNA"
                     /note="constructed by engineering several single-base-pair 
                     substitutions into Filamentous phage cloning vector f8-1 
                     deposited in GenBank Accession number AF218734; vector is 
                     intended as a phage-display vector and has PstI, BamHI, 
                     NheI and MluI cloning sites in gene VIII and confers 
                     tetracycline resistance to the host Escherichia coli cells;
                     'Stiffers' between these cloning sites can be replaced with
                     degenerate coding sequences to create libraries of phage 
                     displaying different random peptides in exposed area of all
                     4000 copies of the major coat protein pVIII"
                     /db_xref="taxon:184976"
     CDS             496..831
                     /codon_start=1
                     /product="replication protein pX"
                     /label=replication protein pX
                     /protein_id="AAM73884.1"
                     /translation="MNIYDDSAVLDAIQSKHFTITPSGKTSFAKASRYFGFYRRLVNEG
                     YDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRPDWYN
                     EPVLKIA"
     CDS             843..1106
                     /codon_start=1
                     /product="cytosolic single-stranded DNA binding protein pV"
                     /label=cytosolic single-stranded DNA binding protein pV
                     /protein_id="AAM73885.1"
                     /translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQLCYVDLGNEYPVLV
                     KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
     CDS             1108..1209
                     /codon_start=1
                     /product="minor coat protein pVII"
                     /label=minor coat protein pVII
                     /protein_id="AAM73886.1"
                     /translation="MEQVADFDTIYQAMIQISVVLCFALGIIAGGQR"
     CDS             1206..1304
                     /codon_start=1
                     /product="minor coat protein pIX"
                     /label=minor coat protein pIX
                     /protein_id="AAM73887.1"
                     /translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
     CDS             1301..1522
                     /codon_start=1
                     /product="major coat protein pVIII precursor"
                     /label=major coat protein pVIII precursor
                     /protein_id="AAM73888.1"
                     /translation="MKKSLVLKASVAVATLVPMLSFAAEGEDPAKAAFDSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     sig_peptide     1301..1369
     terminator      1526..1574
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             1579..2853
                     /codon_start=1
                     /product="adsorption protein pIII precursor"
                     /label=adsorption protein pIII precursor
                     /protein_id="AAM73889.1"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKPHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPYVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     sig_peptide     1579..1632
     CDS             2856..3194
                     /codon_start=1
                     /product="minor coat protein pVI"
                     /label=minor coat protein pVI
                     /protein_id="AAM73890.1"
                     /translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
                     LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKAAIFIFDVKQKI
                     VSYLDWDK"
     CDS             3197..4243
                     /codon_start=1
                     /product="morphogenetic protein pI"
                     /label=morphogenetic protein pI
                     /protein_id="AAM73891.1"
                     /translation="MAVYFVTGKLGSGKTLVSVGKIQDKIVAGCKIATNLDLRLQNLPQ
                     VGRFAKTPRVLRIPDKPSISDLLAIGRGNDSYDENKNGLLVLDECGTWFNTRSWNDKER
                     QPIIDWFLHARKLGWDIIFLVQDLSIVDKQARSALAEHVVYCRRLDRITLPFVGTLYSL
                     VTGSKMPLPKLHVGVVKYGDSQLSPTVERWLYTGKNLYNAYDTKQAFSSNYDSGVYSYL
                     TPYLSHGRYFKPLNLGQKMKLTKIYLKKFSRVLCLAIGFASAFTYSYITQPKPEVKKVV
                     SQTYDFDKFTIDSSQRLNLSYRYVFKDSKGKLINSDDLQKQGYSITYIDLCTVSIKKGN
                     SNEIVKCN"
     CDS             4221..5501
                     /codon_start=1
                     /product="morphogenetic protein IV"
                     /label=morphogenetic protein IV
                     /protein_id="AAM73892.1"
                     /translation="MKLLNVINFVFLMFVSSSSFAQVIEMNNSPLRDFVTWYSKQTGES
                     VIVSPDVKGTVTVYSSDVKPENLRNFFISVLRANNFDMVGSIPSIIQKYNPNSQDYIDE
                     LPSSDIQEYDDNSAPSGGFFVPQNDNVTQTFKINNVRAKDLIRVVELFVKSNTSKSSNV
                     LSVDGSNLLVVSAPKDILDNLPQFLSTVDLPTDQILIEGLIFEVQQGDALDFSFAAGSQ
                     RGTVAGGVNTDRLTSVLSSAGGSFGIFNGDVLGLSVRALKTNSHSKILSVPRILTLSGQ
                     KGSISVGQNVPFITGRVTGESANVNNPFQTVERQNVGISMSVFPVAMAGGNIVLDITSK
                     ADSLSSSTQASDVITNQRSIATTVNLRDGQTLLLGGLTDYKNTSQDSGVPFLSKIPLIG
                     LLFSSRSDSNEESTLYVLVKATIVRAL"
     CDS             complement(6239..7444)
                     /codon_start=1
                     /product="Tn10 tetracycline resistance protein TetA"
                     /label=Tn10 tetracycline resistance protein TetA
                     /protein_id="AAM73882.1"
                     /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
                     /product="Tn10 tetracycline repressor protein TetR"
                     /label=Tn10 tetracycline repressor protein TetR
                     /protein_id="AAM73893.1"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     CDS             8782..831
                     /codon_start=1
                     /product="replication protein pII"
                     /label=replication protein pII
                     /protein_id="AAM73883.1"
                     /translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGMNLSAR
                     TVEYHIDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
                     FGTTDLALCSEALLLNFANSLPCLYDLLDVNATTISRIDATFSARAPNENIAKQVIDHL
                     RNVSNGQTKSTRSQNWESTVTWNETSRHRTLVAYLKHVELQHQIQQLSSKPSAKMTSYQ
                     KEQLKVLSNPDLLEFASGLVRFEARIETRYLKSFGLPLNLFDAIRFASDYNRQGKDLIF
                     DLWSFSFSELFKAFEGDSMNIYDDSAVLDAIQSKHFTITPSGKTSFAKASRYFGFYRRL
                     VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
                     DWYNEPVLKIA"

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