LOCUS       Exported               10832 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCXSN-HA, complete sequence.
ACCESSION   FJ905218
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10832)
  AUTHORS   Chen S, Songkumarn P, Liu J, Wang GL.
  TITLE     A versatile zero background T-vector system for gene cloning and 
            functional genomics
  JOURNAL   Plant Physiol. 150 (3), 1111-1121 (2009)
  PUBMED    19403729
REFERENCE   2  (bases 1 to 10832)
  AUTHORS   Chen S, Songkumarn P, Liu J, Wang G-L.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-APR-2009) Department of Plant Pathology, The Ohio 
            State University, 201 Kottman Hall, 2021 Coffey Rd, Columbus, OH 
            43210, USA
REFERENCE   3  (bases 1 to 10832)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10832
                     /organism="Binary vector pCXSN-HA"
                     /mol_type="other DNA"
                     /db_xref="taxon:643586"
     regulatory      complement(7..270)
                     /regulatory_class="terminator"
                     /note="Nos terminator"
     terminator      8..260
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             627..932
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /protein_id="ACR08768.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            627..932
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    complement(988..1017)
                     /label=HA-tag
                     /note="HA-tag"
     CDS             complement(988..1014)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      complement(1032..1922)
                     /regulatory_class="promoter"
                     /note="CaMV35S promoter"
     promoter        complement(1167..1511)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1937..1953)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2156..2180
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    2174..2199
                     /label=right border T-DNA repeat
                     /note="right border T-DNA repeat"
     misc_feature    complement(3239..4239)
                     /label=pVS1-STA
                     /note="pVS1-STA"
     CDS             3479..4108
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4542..5609
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      complement(4832..5832)
                     /direction=LEFT
                     /label=pVS1-REP
                     /note="pVS1-REP"
     rep_origin      5675..5869
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    6213..6353
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    complement(6242..6502)
                     /label=pBR-BOM
                     /note="pBR-BOM"
     rep_origin      complement(6539..7127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(6642..6922)
                     /direction=LEFT
                     /label=pBR-ORI
                     /note="pBR-ORI"
     CDS             complement(7214..8008)
                     /codon_start=1
                     /gene="aadA"
                     /product="kanamycin resistance protein"
                     /label=aadA
                     /protein_id="ACR08766.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(7214..8008)
                     /gene="aadA"
                     /label=aadA
     misc_feature    8432..8457
                     /label=left border T-DNA repeat
                     /note="left border T-DNA repeat"
     misc_feature    8433..8457
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(8524..8732)
                     /label=CaMV35S polyA signal
                     /note="CaMV35S polyA signal"
     polyA_signal    8535..8709
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(8749..9774)
                     /codon_start=1
                     /gene="hptII"
                     /product="hygromycin resistance protein"
                     /label=hptII
                     /note="hygromycin phosphotransferase"
                     /protein_id="ACR08767.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            complement(8749..9774)
                     /gene="hptII"
                     /label=hptII
     regulatory      complement(9808..10588)
                     /regulatory_class="promoter"
                     /note="CaMV35S promoter"
     promoter        complement(9841..10518)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    10709..10730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        10745..10775
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10783..10799
                     /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)."
     primer_bind     10807..10823
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"