LOCUS       Exported               11891 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPOP6IPT, complete sequence.
ACCESSION   JX185749
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11891)
  AUTHORS   Schafer M, Brutting C, Gase K, Reichelt M, Baldwin I, Meldau S.
  TITLE     Real time genetic manipulation: a new tool for ecological field 
            studies
  JOURNAL   Plant J. 76 (3), 506-518 (2013)
  PUBMED    23906159
REFERENCE   2  (bases 1 to 11891)
  AUTHORS   Gase K, Meldau S, Schaefer M, Baldwin IT.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-JUN-2012) Molecular Ecology, Max-Planck-Institute for 
            Chemical Ecology, Hans-Knoell-Strasse 8, Jena, Thueringen 07745, 
            Germany
REFERENCE   3  (bases 1 to 11891)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11891
                     /organism="Cloning vector pPOP6IPT"
                     /mol_type="other DNA"
                     /db_xref="taxon:1307966"
     regulatory      complement(1..345)
                     /regulatory_class="terminator"
                     /note="Agrobacterium tumefaciens isopentenyl 
                     phosphotransferase"
     CDS             complement(417..1442)
                     /codon_start=1
                     /product="Hpt"
                     /label=Hpt
                     /note="hygromycin phosphotransferase"
                     /protein_id="AGI38106.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     regulatory      complement(1469..1756)
                     /regulatory_class="promoter"
                     /note="NOS"
     promoter        complement(1494..1673)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     complement(2094..2110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2118..2134
                     /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(2142..2172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2187..2208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2435..3194
                     /label=ColE1 type from pUC19
                     /note="ColE1 type from pUC19"
     rep_origin      complement(2496..3084)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3368..4162)
                     /codon_start=1
                     /product="NptII"
                     /label=NptII
                     /note="neomycin phosphotransferase II"
                     /protein_id="AGI38107.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      complement(4266..4271)
                     /regulatory_class="minus_10_signal"
                     /note="Transposon Tn5"
     regulatory      complement(4289..4294)
                     /regulatory_class="minus_35_signal"
                     /note="Transposon Tn5"
     primer_bind     4667..4683
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_structure  4729..5082
                     /note="pOp6 promoter from: Samalova, M. et al. (2005) 
                     pOp6/LhGR: A stringently regulated and highly responsive 
                     dexamethasone-inducible gene expression system for tobacco.
                     The Plant Journal 41: 919-935"
     misc_feature    join(4729..4748,4781..4800,4833..4852,4885..4904,4937..4956,
                     4989..5008)
                     /note="AATTGTGAGCGCTCACAATT = ideal lac operator; from: 
                     Lehming et al. (1987) The interaction of the recognition 
                     helix opf lac repressor with lac operator. EMBO J. 6: 
                     3145-3153"
     protein_bind    4729..4748
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    4781..4800
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    4833..4852
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    4885..4904
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    4937..4956
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    4989..5008
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     regulatory      5028..5082
                     /regulatory_class="promoter"
                     /note="cauliflower mosaic virus minimal 35S"
     promoter        5032..5078
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             5111..5833
                     /codon_start=1
                     /product="IPT"
                     /label=IPT
                     /note="Agrobacterium tumefaciens isopentenyl transferase"
                     /protein_id="AGI38108.1"
                     /translation="MDLHLIFGPTCTGKTTTAIALAQQTGLPVLSLDRVQCCPQLSTGS
                     GRPTVEELKGTTRLYLDDRPLVEGIIAAKQAHHRLIEEVYNHEANGGLILEGGSTSLLN
                     CMARNSYWSADFRWHIIRHKLPDQETFMKAAKARVKQMLHPAAGHSIIQELVYLWNEPR
                     LRPILKEIDGYRYAMLFASQNQITADMLLQLDANMEGKLINGIAQEYFIHARQQEQKFP
                     QVNAAAFDGFEGHPFGMY"
     regulatory      5899..6058
                     /regulatory_class="terminator"
                     /note="cauliflower mosaic virus"
     misc_feature    6124..6149
                     /label=right border T-DNA
                     /note="right border T-DNA"
     misc_feature    6125..6149
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      7303..7497
                     /label=pVS1
                     /note="pVS1"
     CDS             complement(7563..8738)
                     /codon_start=1
                     /product="pVS1 RepA"
                     /label=pVS1 RepA
                     /note="pVS1 replication protein"
                     /protein_id="AGI38109.1"
                     /translation="MPHVWRNGRLARRKRLGCLPALQWHWNPQARGIGVSGRKPSGPVQ
                     IGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQAAADRIRKESRQPPAAGAPSIRKP
                     PKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKRDRRAGEVIRYELPDGHVEVSAGP
                     AGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGRVFRPHVADVLKFCRRADGGKQKD
                     DLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEALISRYKIVKSETGRPEYIEIELAD
                     WMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLARRAAGKAEARWLFKTIYERSGSA
                     GEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILVMRYRNLIEGEASAGS"
     CDS             complement(7563..8636)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(9065..9694)
                     /codon_start=1
                     /product="pVS1 StaA"
                     /label=pVS1 StaA
                     /note="pVS1 partitioning protein"
                     /protein_id="AGI38110.1"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    11178..11202
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    11178..11200
                     /label=left border T-DNA
                     /note="left border T-DNA"