LOCUS       Exported               12827 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Promoter probe vector pRR21, complete sequence.
ACCESSION   EU760495
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12827)
  AUTHORS   Raveendra GM, Bhat RS, Kuruvinashetti MS.
  TITLE     Promoter probe vector with Sgfp65T reporter
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12827)
  AUTHORS   Raveendra GM, Bhat RS, Kuruvinashetti MS.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-MAY-2008) Department of Biotechnology, University of 
            Agricultural Sciences, Krishinagar, Dharwad, Karnataka 580 005, 
            India
REFERENCE   3  (bases 1 to 12827)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12827
                     /organism="Promoter probe vector pRR21"
                     /mol_type="other DNA"
                     /db_xref="taxon:542345"
     terminator      7..259
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      complement(270..12827)
                     /regulatory_class="terminator"
                     /note="NosT"
     CDS             complement(282..1001)
                     /codon_start=1
                     /gene="Sgfp65T"
                     /product="Sgfp65T"
                     /label=Sgfp65T
                     /note="synthetic GFP"
                     /protein_id="ACE95705.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     gene            complement(282..1001)
                     /gene="Sgfp65T"
                     /label=Sgfp65T
     primer_bind     complement(1041..1057)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1251..1788
                     /regulatory_class="promoter"
                     /note="CaMV35"
     promoter        1434..1779
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             join(1796..1810,2001..3848)
                     /codon_start=1
                     /gene="GUSPlus-His6"
                     /product="GUSPlus-His6"
                     /label=GUSPlus-His6
                     /note="GUS reporter"
                     /protein_id="ACE95706.1"
                     /translation="MVDLRNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYES
                     KLTDTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAI
                     VYVNGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGL
                     GKVIRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAE
                     TVKVSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEP
                     FGVRTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFR
                     TAHYPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHH
                     QDVLRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLF
                     VMATPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITE
                     YGADTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRV
                     QGNKKGVFTRDRKPKLAAHVFRERWTNIPDFGYKNASHHHHHHV"
     gene            1796..3848
                     /gene="GUSPlus-His6"
                     /label=GUSPlus-His6
     CDS             join(1796..1810,2001..3818)
                     /codon_start=1
                     /product="beta-glucuronidase"
                     /label=GUSPlus(TM)
                     /note="codon-optimized Staphylococcus gusA gene with a 
                     catalase intron to ensure expression in plants but not 
                     bacteria"
                     /translation="MVDLRNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYES
                     KLTDTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAI
                     VYVNGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGL
                     GKVIRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAE
                     TVKVSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEP
                     FGVRTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFR
                     TAHYPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHH
                     QDVLRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLF
                     VMATPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITE
                     YGADTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRV
                     QGNKKGVFTRDRKPKLAAHVFRERWTNIPDFGYKN"
     intron          1811..2000
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     CDS             3825..3842
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3874..4126
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3877..4129
                     /label=polyA signal
                     /note="polyA signal"
     misc_feature    4136..4172
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    4148..4172
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             5472..6101
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6535..7602
                     /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      6826..7826
     rep_origin      7668..7862
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    8206..8346
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(8532..9120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      8636..8919
                     /label=pBR322 ori
                     /note="pBR322 ori"
     CDS             complement(9207..10001)
                     /codon_start=1
                     /gene="Kan"
                     /product="neomycin phosphotransferase"
                     /label=Kan
                     /note="confers resistance to kanamycin"
                     /protein_id="ACE95707.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(9207..10001)
                     /gene="Kan"
                     /label=Kan
     misc_feature    complement(10426..10470)
                     /label=T-DNA left border
                     /note="T-DNA left border"
     misc_feature    10426..10450
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    10528..10702
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     regulatory      complement(10592..10599)
                     /regulatory_class="polyA_signal_sequence"
     CDS             complement(10742..11767)
                     /codon_start=1
                     /gene="hpt"
                     /product="hygromycin phosphotransferase"
                     /label=hpt
                     /protein_id="ACE95708.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            complement(10742..11767)
                     /gene="hpt"
                     /label=hpt
     regulatory      complement(11803..12610)
                     /regulatory_class="promoter"
                     /note="CaMV35S"
     promoter        complement(11835..12512)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    12703..12724
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        12739..12769
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12777..12793
                     /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     12801..12817
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"