LOCUS       Exported               10480 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCLEAN-S167, complete sequence.
ACCESSION   EU186086
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10480)
  AUTHORS   Thole V, Worland B, Snape JW, Vain P.
  TITLE     The pCLEAN dual binary vector system for Agrobacterium-mediated 
            plant transformation
  JOURNAL   Plant Physiol. 145 (4), 1211-1219 (2007)
  PUBMED    17932303
REFERENCE   2  (bases 1 to 10480)
  AUTHORS   Thole V, Vain P.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-SEP-2007) Department of Crop Genetics, John Innes 
            Centre, Colney Lane, Norwich NR4 7UH, UK
REFERENCE   3  (bases 1 to 10480)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10480
                     /organism="Binary vector pCLEAN-S167"
                     /mol_type="other DNA"
                     /db_xref="taxon:478697"
     misc_feature    1..9224
                     /note="Cloning vector pSoup (Genbank Accession Number 
                     EU048870)-based backbone sequence"
     promoter        complement(7..25)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(43..59)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    67..83
                     /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(91..121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    136..157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        404..432
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             480..1670
                     /codon_start=1
                     /gene="tetA"
                     /product="TetA"
                     /label=tetA
                     /note="tetracycline resistance protein class C"
                     /protein_id="ABW84169.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            480..1670
                     /gene="tetA"
                     /label=tetA
     CDS             2169..3317
                     /codon_start=1
                     /gene="trfA"
                     /product="TrfA"
                     /label=trfA
                     /protein_id="ABW84170.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            2169..3317
                     /gene="trfA"
                     /label=trfA
     CDS             4698..5669
                     /codon_start=1
                     /gene="pSa Rep"
                     /product="pSa Rep"
                     /label=pSa Rep
                     /note="pSa replicase; pSa-ori is located on other binary 
                     vectors such as pGreen and pCLEAN-G"
                     /protein_id="ABW84171.1"
                     /translation="MPKNNKAPGHRINEIIKTSLALEMEDAREAGLVGYMARCLVQATM
                     PHTDPKTSYFERTNGIVTLSIMGKPSIGLPYGSMPRTLLAWICTEAVRTKDPVLNLGRS
                     QSEFLQRLGMHTDGRYTATLRNQAQRLFSSMISLAGEQGNDFGIENVVIAKRAFLFWNP
                     KRPEDRALWDSTLTLTGDFFEEVTRSPVPIRIDYLHALRQSPLAMDIYTWLTYRVFLLR
                     AKGRPFVQIPWVALQAQFGSSYGSRARNSPELDDKARERAERAALASFKYNFKKRLREV
                     LIVYPEASDCIEDDGECLRIKSTRLHVTRAPGKGARIGPPPT"
     gene            4698..5669
                     /gene="pSa Rep"
                     /label=pSa Rep
     CDS             6064..6714
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     rep_origin      7013..7871
                     /label=ColE1-ori
                     /note="ColE1-ori"
     oriT            7395..7504
                     /note="incP origin of transfer"
     CDS             7537..7908
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      8035..8746
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      8053..8670
                     /label=OriV
                     /note="OriV"
     misc_feature    9225..10480
                     /note="Basic pCLEAN T-DNA region no. 8 including minimal 
                     T-DNA of 102 nucleotides (Thole et al., 2007; 
                     http://www.jic.ac.uk/staff/philippe-vain/vectors.htm)"
     misc_feature    9246..9270
                     /note="25-nt long consensus T-DNA left border sequence of 
                     the nopaline vector pTiT37 (Slightom et al., (1985) EMBO J 
                     4:3069-3077)"
     misc_feature    9271..9322
                     /note="Inner T-DNA region containing 52 nt-long multiple 
                     cloning site with NotI, PacI, SfiI, BsiWI, ApaI, XhoI and 
                     SalI restriction sites"
     misc_feature    9323..9347
                     /note="25-nt long consensus T-DNA right border sequence of 
                     the nopaline vector pTiT37 (Slightom et al., (1985) EMBO J 
                     4:3069-3077)"
     misc_feature    9348..9371
                     /note="right border overdrive sequence (Peralta et al. 
                     (1986) EMBO J 5: 1137-1142)"
     CDS             9585..10388
                     /codon_start=1
                     /gene="virGwt"
                     /product="VirGwt"
                     /label=virGwt
                     /note="wild-type virG"
                     /protein_id="ABW84172.1"
                     /translation="MIVHPSRENFSSAVNKGSDFRLKGEPLKHVLLVDDDVAMRHLIIE
                     YLTIHAFKVTAVADSTQFTRVLSSATVDVVVVDLNLGREDGLEIVRNLAAKSDIPIIII
                     SGDRLEETDKVVALELGASDFIAKPFSIREFLARIRVALRVRPNVVRSKDRRSFCFTDW
                     TLNLRQRRLMSEAGGEVKLTAGEFNLLLAFLEKPRDVLSREQLLIASRVRDEEVYDRSI
                     DVLILRLRRKLEADPSSPQLIKTARGAGYFFDADVQVSHGGTMAA"
     gene            9585..10388
                     /gene="virGwt"
                     /label=virGwt