LOCUS       Exported               12189 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Gateway compatible binary vector pCS167, complete sequence.
ACCESSION   KT163018
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12189)
  AUTHORS   Wang G, Jones H, Doherty A, Yu X, Ma Y, Sun Y, Du W, Wang C, Xia L.
  TITLE     Generation of marker- and backbone-free transgenic wheat plants via 
            Agrobacterium-mediated transformation based on the pCLEAN dual 
            binary vector system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12189)
  AUTHORS   Wang G, Jones H, Doherty A, Yu X, Ma Y, Sun Y, Du W, Wang C, Xia L.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JUN-2015) Institute of Crop Sciences, Chinese Academy 
            of Agricultural Sciences, 12 Zhongguancun Street, Beijing, Beijing 
            100081, China
REFERENCE   3  (bases 1 to 12189)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..12189
                     /organism="Gateway compatible binary vector pCS167"
                     /mol_type="other DNA"
                     /db_xref="taxon:1739643"
     misc_feature    1..9224
                     /note="Cloning vector pSoup (INSD 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="ALK43910.1"
                     /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="ALK43911.1"
                     /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="TrfA"
                     /label=pSa Rep
                     /note="pSa replicase"
                     /protein_id="ALK43912.1"
                     /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    9246..9270
                     /note="T-DNA left border sequence (Thole et al., (2007) 
                     Plant Physiol 145:1211-1219)"
     misc_recomb     9291..9415
                     /note="attR2; gateway recombination site; derived from 
                     Escherichia coli"
     protein_bind    9291..9415
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(9456..9761)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(9465..9761)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /note="cell cycle death B; cytotoxic protein derived from 
                     Escherichia coli"
                     /protein_id="ALK43913.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFW"
     gene            complement(9465..9761)
                     /gene="ccdB"
                     /label=ccdB
     CDS             complement(10103..10762)
                     /codon_start=1
                     /gene="cmR"
                     /product="CmR"
                     /label=cmR
                     /note="chloramphenicol resistance protein"
                     /protein_id="ALK43914.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(10103..10762)
                     /gene="cmR"
                     /label=cmR
     promoter        complement(10816..10846)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_recomb     10871..10995
                     /note="attR1; gateway recombination site; derived from 
                     Escherichia coli"
     protein_bind    complement(10871..10995)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    11032..11056
                     /note="T-DNA right border sequence (Thole et al., (2007) 
                     Plant Physiol 145:1211-1219)"
     misc_feature    11057..11080
                     /note="right border overdrive sequence (Thole et al., 
                     (2007) Plant Physiol 145:1211-1219)"
     CDS             11294..12097
                     /codon_start=1
                     /gene="virGwt"
                     /product="VirGwt"
                     /label=virGwt
                     /note="wild-type virG"
                     /protein_id="ALK43915.1"
                     /translation="MIVHPSRENFSSAVNKGSDFRLKGEPLKHVLLVDDDVAMRHLIIE
                     YLTIHAFKVTAVADSTQFTRVLSSATVDVVVVDLNLGREDGLEIVRNLAAKSDIPIIII
                     SGDRLEETDKVVALELGASDFIAKPFSIREFLARIRVALRVRPNVVRSKDRRSFCFTDW
                     TLNLRQRRLMSEAGGEVKLTAGEFNLLLAFLEKPRDVLSREQLLIASRVRDEEVYDRSI
                     DVLILRLRRKLEADPSSPQLIKTARGAGYFFDADVQVSHGGTMAA"
     gene            11294..12097
                     /gene="virGwt"
                     /label=virGwt