fd-tet vector (V009986)

Basic Vector Information

      • Vector Name:
      • fd-tet
      • Antibiotic Resistance:
      • Tetracycline
      • Length:
      • 9183 bp
      • Type:
      • Filamentous phage cloning vector
      • Source/Author:
      • Beck E, Sommer R, Auerswald EA, Kurz C, Zink B, Osterburg G, Schaller H, Sugimoto K, Sugisaki H, Okamoto T, Takanami M.

fd-tet vector Vector Map

fd-tet9183 bp40080012001600200024002800320036004000440048005200560060006400680072007600800084008800replication protein pXcytosolic single-stranded DNA binding protein pVminor coat protein pVIImajor coat protein pVIII precursorfd terminatoradsorption protein pIII precursorminor coat protein pVImorphogenetic protein pImorphogenetic protein signal sequence; promotes packaging of circular single plus strands into progeny virions; other siteminus-strand origin, disrupted by Tn10-derived insert; functionally inactive; minus strand initiated randomly by endogenous bacterial Okazaki fragment priming apparatusplus strand origin

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 ℃.

fd-tet vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                9183 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Filamentous phage cloning vector fd-tet, complete sequence.
ACCESSION   AF217317
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9183)
  AUTHORS   Beck E, Sommer R, Auerswald EA, Kurz C, Zink B, Osterburg G, 
            Schaller H, Sugimoto K, Sugisaki H, Okamoto T, Takanami M.
  TITLE     Nucleotide sequence of bacteriophage fd DNA
  JOURNAL   Nucleic Acids Res. 5 (12), 4495-4503 (1978)
  PUBMED    745987
REFERENCE   2  (bases 1 to 9183)
  AUTHORS   Zacher AN III, Stock CA, Golden JW II, Smith GP.
  TITLE     A new filamentous phage cloning vector: fd-tet
  JOURNAL   Gene 9 (1-2), 127-140 (1980)
  PUBMED    6247241
REFERENCE   3  (bases 1 to 9183)
  AUTHORS   Zinder ND, Horiuchi K.
  TITLE     Multiregulatory element of filamentous bacteriophages
  JOURNAL   Microbiol. Rev. 49 (2), 101-106 (1985)
  PUBMED    2989671
REFERENCE   4  (bases 1 to 9183)
  AUTHORS   Smith GP.
  TITLE     Filamentous phage assembly: morphogenetically defective mutants that
            do not kill the host
  JOURNAL   Virology 167 (1), 156-165 (1988)
  PUBMED    3188394
REFERENCE   5  (bases 1 to 9183)
  AUTHORS   Smith GP.
  TITLE     Filamentous phage vector fd-tet
  JOURNAL   Unpublished
REFERENCE   6  (bases 1 to 9183)
  AUTHORS   Smith GP.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-DEC-1999) Division of Biological Sciences, University 
            of Missouri, Tucker Hall, Columbia, MO 65211-7400, USA
REFERENCE   7  (bases 1 to 9183)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     Filamentous phage cloning vector constructed by  splicing the 
            tetracycline resistance determinant of Tn10 into the minus-strand 
            origin of bacteriphage fd.  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.  Transduce tetracycline resistance into infected 
            host cell.  Make  very small plaques.  Foreign inserts can be 
            spliced into unique HindIII site, and are generally stable.
FEATURES             Location/Qualifiers
     source          1..9183
                     /organism="Filamentous phage cloning vector fd-tet"
                     /focus
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:127516"
     source          1..5645
                     /organism="Enterobacteria phage fd"
                     /mol_type="other DNA"
                     /db_xref="taxon:10864"
     source          8421..9183
                     /organism="Enterobacteria phage fd"
                     /mol_type="other DNA"
                     /db_xref="taxon:10864"
     CDS             496..831
                     /codon_start=1
                     /product="replication protein pX"
                     /label=replication protein pX
                     /protein_id="AAF75601.1"
                     /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="AAF75603.1"
                     /translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQLCYVDLGNEYPVLV
                     KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
     CDS             1108..1209
                     /codon_start=1
                     /product="minor coat protein pVII"
                     /label=minor coat protein pVII
                     /protein_id="AAF75604.1"
                     /translation="MEQVADFDTIYQAMIQISVVLCFALGIIAGGQR"
     CDS             1206..1304
                     /codon_start=1
                     /product="minor coat protein pIX"
                     /label=minor coat protein pIX
                     /protein_id="AAF75605.1"
                     /translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
     CDS             1301..1522
                     /codon_start=1
                     /product="major coat protein pVIII precursor"
                     /label=major coat protein pVIII precursor
                     /protein_id="AAF75598.1"
                     /translation="MKKSLVLKASVAVATLVPMLSFAAEGDDPAKAAFDSLQASATEYI
                     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="AAF75609.1"
                     /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="AAF75606.1"
                     /translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
                     LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKAAIFIFDVKQKI
                     VSYLDWDK"
     CDS             3197..4243
                     /codon_start=1
                     /product="morphogenetic protein pI"
                     /label=morphogenetic protein pI
                     /protein_id="AAF75599.1"
                     /translation="MAVYFVTGKLGSGKTLVSVGKIQDKIVAGCKIATNLDLRLQNLPQ
                     VGRFAKTPRVLRIPDKPSISDLLAIGRGNDSYDENKNGLLVLDECGTWFNTRSWNDKER
                     QPIIDWFLHARKLGWDIIFLVQDLSIVDKQARSALAEHVVYCRRLDRITLPFVGTLYSL
                     VTGSKMPLPKLHVGVVKYGDSQLSPTVERWLYTGKNLYNAYDTKQAFSSNYDSGVYSYL
                     TPYLSHGRYFKPLNLGQKMKLTKIYLKKFSRVLCLAIGFASAFTYSYITQPKPEVKKVV
                     SQTYDFDKFTIDSSQRLNLSYRYVFKDSKGKLINSDDLQKQGYSITYIDLCTVSIKKGN
                     SNEIVKCN"
     CDS             4221..5501
                     /codon_start=1
                     /product="morphogenetic protein IV"
                     /label=morphogenetic protein IV
                     /protein_id="AAF75602.1"
                     /translation="MKLLNVINFVFLMFVSSSSFAQVIEMNNSPLRDFVTWYSKQTGES
                     VIVSPDVKGTVTVYSSDVKPENLRNFFISVLRANNFDMVGSIPSIIQKYNPNSQDYIDE
                     LPSSDIQEYDDNSAPSGGFFVPQNDNVTQTFKINNVRAKDLIRVVELFVKSNTSKSSNV
                     LSVDGSNLLVVSAPKDILDNLPQFLSTVDLPTDQILIEGLIFEVQQGDALDFSFAAGSQ
                     RGTVAGGVNTDRLTSVLSSAGGSFGIFNGDVLGLSVRALKTNSHSKILSVPRILTLSGQ
                     KGSISVGQNVPFITGRVTGESANVNNPFQTVERQNVGISMSVFPVAMAGGNIVLDITSK
                     ADSLSSSTQASDVITNQRSIATTVNLRDGQTLLLGGLTDYKNTSQDSGVPFLSKIPLIG
                     LLFSSRSDSNEESTLYVLVKATIVRAL"
     misc_feature    5500..5577
                     /note="morphogenetic protein signal sequence; promotes 
                     packaging of circular single plus strands into progeny 
                     virions; other site"
                     /citation="[3]"
     rep_origin      join(5624..5645,8421..8532)
                     /note="minus-strand origin, disrupted by Tn10-derived 
                     insert; functionally inactive; minus strand initiated 
                     randomly by endogenous bacterial Okazaki fragment priming 
                     apparatus"
                     /citation="[3]"
                     /citation="[4]"
     mobile_element  5646..8420
                     /mobile_element_type="transposon:Tn10"
     misc_feature    5975..5980
                     /label=HindIII cloning site
                     /note="HindIII cloning site"
     CDS             complement(6239..7444)
                     /codon_start=1
                     /product="Tn10 tetracycline resistance protein TetA"
                     /label=Tn10 tetracycline resistance protein TetA
                     /protein_id="AAF75607.1"
                     /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="AAF75608.1"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     rep_origin      8541..8680
                     /label=plus strand origin
                     /note="plus strand origin"
                     /citation="[3]"
     misc_feature    8556..8557
                     /note="pII replication protein nicks plus strand to 
                     initiate rolling circle replication"
     misc_difference 8771
                     /replace="c"
                     /citation="[1]"
     CDS             8782..831
                     /codon_start=1
                     /product="replication protein pII"
                     /label=replication protein pII
                     /protein_id="AAF75600.1"
                     /translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGMNLSAR
                     TVEYHIDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
                     FGTTDLALCSEALLLNFANSLPCLYDLLDVNATTISRIDATFSARAPNENIAKQVIDHL
                     RNVSNGQTKSTRSQNWESTVTWNETSRHRTLVAYLKHVELQHQIQQLSSKPSAKMTSYQ
                     KEQLKVLSNPDLLEFASGLVRFEARIETRYLKSFGLPLNLFDAIRFASDYNRQGKDLIF
                     DLWSFSFSELFKAFEGDSMNIYDDSAVLDAIQSKHFTITPSGKTSFAKASRYFGFYRRL
                     VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
                     DWYNEPVLKIA"

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