LOCUS       Exported               13972 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pHZPRi-Hyg, complete sequence.
ACCESSION   HM480107
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13972)
  AUTHORS   Zhang H, Timko MP.
  TITLE     The tobacco transcription factors NtMYC2a and NtMYC2b regulate 
            multiple steps in nicotine biosynthesis and form nuclear complexes 
            with NtJAZ1 repressor
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13972)
  AUTHORS   Zhang H, Timko MP.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JUN-2010) Biology, University of Virginia, Department 
            of Biology, University of Virginia, Charlottesville, VA 22904, USA
REFERENCE   3  (bases 1 to 13972)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13972
                     /organism="Binary vector pHZPRi-Hyg"
                     /mol_type="other DNA"
                     /db_xref="taxon:941634"
     regulatory      1..260
                     /regulatory_class="terminator"
                     /note="Nos terminator"
     misc_recomb     301..425
                     /label=attR1 gateway recombination site
                     /note="attR1 gateway recombination site"
     protein_bind    301..425
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        450..480
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             534..1193
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=chloramphenicol acetyltransferase
                     /note="confers resistance to antibiotic chloramphenicol"
                     /protein_id="ADU87116.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1535..1840
                     /codon_start=1
                     /product="cytotoxic protein CcdB"
                     /label=cytotoxic protein CcdB
                     /protein_id="ADU87117.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_recomb     1881..2005
                     /label=attR2 gateway recombination site
                     /note="attR2 gateway recombination site"
     protein_bind    complement(1881..2005)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2017..2425
                     /label=partial GUS fragment
                     /note="partial GUS fragment"
     misc_recomb     2441..2565
                     /label=attR2 gateway recombination site
                     /note="attR2 gateway recombination site"
     protein_bind    2441..2565
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2606..2911)
                     /codon_start=1
                     /product="cytotoxic protein CcdB"
                     /label=cytotoxic protein CcdB
                     /protein_id="ADU87118.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(3253..3912)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=chloramphenicol acetyltransferase
                     /note="confers resistance to antibiotic chloramphenicol"
                     /protein_id="ADU87119.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3966..3996)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_recomb     complement(4021..4145)
                     /label=attR1 gateway recombination site
                     /note="attR1 gateway recombination site"
     protein_bind    complement(4021..4145)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(4298..4642)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     regulatory      4299..5053
                     /regulatory_class="promoter"
                     /note="2X CaMV 35S promoter"
     primer_bind     complement(5074..5090)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5293..5318
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    5293..5317
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6617..7246
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             7680..8747
                     /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      8813..9007
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    9351..9491
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(9677..10265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10352..11146)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=aminoglycoside phosphotransferase
                     /note="confers resistance to antibiotic kanamycin in 
                     bacterial"
                     /protein_id="ADU87120.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    11570..11595
                     /label=T-DNA left border
                     /note="T-DNA left border"
     misc_feature    11571..11595
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    11673..11847
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(11887..12912)
                     /codon_start=1
                     /product="hygromycin phosphotransferase"
                     /label=hygromycin phosphotransferase
                     /note="confers resistance to antibiotic hygromycin in 
                     plant"
                     /protein_id="ADU87121.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(12980..13657)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    13848..13869
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        13884..13914
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13922..13938
                     /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     13946..13962
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"