pRK404 vector (V003727)

Basic Vector Information

Vector Name:
pRK404
Antibiotic Resistance:
Tetracycline
Length:
11237 bp
Type:
Cloning vector
Replication origin:
oriV
Source/Author:
Scott HN, Laible PD, Hanson DK.

pRK404 vector Map

pRK40411237 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000oriVlacZalac operatorlac promoterCAP binding sitetraJoriTTetRTcRtrfAssbtrbAincC

pRK404 vector Sequence

LOCUS       40924_37118       11237 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRK404, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11237)
  AUTHORS   Scott HN, Laible PD, Hanson DK.
  TITLE     Sequences of versatile broad-host-range vectors of the RK2 family
  JOURNAL   Plasmid 50 (1), 74-79 (2003)
  PUBMED    12826060
REFERENCE   2  (bases 1 to 11237)
  AUTHORS   Scott HN, Laible PD, Hanson DK.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-DEC-2002) Biosciences Division, Argonne National 
            Laboratory, 9700 South Cass Ave., Argonne, IL 60439, USA
REFERENCE   3  (bases 1 to 11237)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11237)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2003"; volume: "50"; issue: "1"; pages: "74-79"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-DEC-2002) Biosciences Division, Argonne National Laboratory, 
            9700 South Cass Ave., Argonne, IL 60439, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(276..987)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     CDS             complement(1246..1566)
                     /codon_start=1
                     /gene="lacZa"
                     /product="beta-galactosidase alpha peptide"
                     /label=lacZa
                     /protein_id="AAO47400.1"
                     /translation="MTMITPSLAAGRRIPGNSLAVVLQRRDWENPGVTQLNRLAAHPPF
                     ASWRNSEEARTDRPSQQLRSLNGEWRHASRREKGGQVSGKRQGRNRRAHEGASRGKRLV
                     SL"
     gene            complement(1246..1566)
                     /gene="lacZa"
                     /label=lacZa
     primer_bind     1497..1513
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1562..1578)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1586..1602
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1610..1640)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1655..1676)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1911..2279)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(2312..2421)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(3105..3752)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             3858..5054
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             complement(6333..7478)
                     /codon_start=1
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(7527..7877)
                     /codon_start=1
                     /gene="ssb"
                     /product="single-stranded DNA binding protein"
                     /label=ssb
                     /protein_id="AAO47405.1"
                     /translation="MSHNQFQFIGNLTRDTEVRHGNSNKPQAIFDIAVNEEWRNDAGDK
                     QERTDFFRIKCFGSQAEAHGKYLGKGSLVFVQGKIRNTKYEKDGQTVYGTDFIADKVDY
                     LDTKAPGGSNQE"
     gene            complement(7527..7877)
                     /gene="ssb"
                     /label=ssb
     CDS             8052..8363
                     /codon_start=1
                     /gene="trbA"
                     /product="regulation of Tra gene expression"
                     /label=trbA
                     /protein_id="AAO47406.1"
                     /translation="MTKHELSERAGVSISFLSDLTNGKANPSLKVMEAIADALETPLPL
                     LLESTDLDREALAEIAGHPFKSSVPPGYERISVVLPSHKAFIVKKWGDDTRKKLRGRL"
     gene            8052..8363
                     /gene="trbA"
                     /label=trbA
     CDS             complement(9401..10495)
                     /codon_start=1
                     /gene="incC"
                     /product="plasmid maintenance protein"
                     /label=incC
                     /protein_id="AAO47407.1"
                     /translation="MGVIHEETAYRKPVPGGDPGAGSGAADHRDSAGRLSRWEATGDVR
                     NVAGTDQGRSVASGASRVGRVRGQELARGVRAGNGGSAGTSGVHRPEVGSGRQEKTGNQ
                     TMKTLVTANQKGGVGKTSTLVHLAFDFFERGLRVAVIDLDPQGNASYTLKDFATGLHAS
                     KLFGAVPAGGWTETAPAAGDGQAARLALIESNPVLANAERLSLDDARELFGANIKALAN
                     QGFDVCLIDTAPTLGVGLAAALFAADYVLSPIELEAYSIQGIKKMVTTIANVRQKNAKL
                     QFLGMVPSKVDARNPRHARHQAELLAAYPKMMIPATVGLRSSIADALASGVPVWKIKKT
                     AARKASKEVRALADYVFTKMEISQ"
     gene            complement(9401..10495)
                     /gene="incC"
                     /label=incC
     CDS             complement(10177..10482)
                     /codon_start=1
                     /gene="korA"
                     /product="regulation of gene expression"
                     /label=korA
                     /protein_id="AAO47408.1"
                     /translation="MKKRLTESQFQEAIQGLEVGQQTIEIARGVLVDGKPQATFATSLG
                     LTRGAVSQAVHRVWAAFEDKNLPEGYARVTAVLPEHQAYIVRKWEADAKKKQETKR"
     gene            complement(10177..10482)
                     /gene="korA"
                     /label=korA

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