LOCUS Exported 12361 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Binary vector pCAMBIA-1303, complete sequence.
ACCESSION AF234299
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12361)
AUTHORS Hajdukiewicz P, Svab Z, Maliga P.
TITLE The small, versatile pPZP family of Agrobacterium binary vectors for
plant transformation
JOURNAL Plant Mol. Biol. 25 (6), 989-994 (1994)
PUBMED 7919218
REFERENCE 2 (bases 1 to 12361)
AUTHORS Roberts C, Rajagopal S, Smith LM, Nguyen TA, Yang W, Nugrohu S, Ravi
KS, Vijayachandra K, Harcourt RL, Dransfield L, Desamero N, Slamet
I, Hadjukiewicz P, Svab Z, Maliga P, Mayer JE, Keese PK, Kilian A,
Jefferson RA.
TITLE A comprehensive set of modular vectors for advanced manipulations
and efficient transformation of plants
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 12361)
AUTHORS Roberts C, Rajagopal S, Smith LM, Nguyen TA, Yang W, Nugrohu S, Ravi
KS, Vijayachandra K, Harcourt RL, Dransfield L, Desamero N, Slamet
I, Hadjukiewicz P, Svab Z, Maliga P, Mayer JE, Keese PK, Kilian A,
Jefferson RA.
TITLE Direct Submission
JOURNAL Submitted (15-FEB-2000) CAMBIA, Clunies Ross St, Black Mountain /
GPO Box 3200, Canberra, ACT 2601, Australia
REFERENCE 4 (bases 1 to 12361)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..12361
/organism="Binary vector pCAMBIA-1303"
/mol_type="other DNA"
/db_xref="taxon:118391"
CDS 2..2569
/codon_start=1
/product="GUSA (N358Q)/MGFP5 fusion, hexaHis tagged"
/label=GUSA
/note="gusA (N358Q) - mgfp5* gene fusion; His6"
/protein_id="AAF65347.1"
/translation="MVDLTSLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQ
ESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWV
NNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQ
SYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELR
DADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAV
KGQQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAE
EMLDWADEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKE
LIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHT
DTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAG
LHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIF
TRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQASKGEELFTGVVPILVELDGDVNGHKF
SVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKS
AMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNY
NSHNVYIMADKQKNGIKANFKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQ
SALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKASHHHHHHV"
CDS 20..1825
/codon_start=1
/gene="uidA (or gusA) from E. coli"
/product="beta-glucuronidase"
/label=GUS
/translation="LRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAIA
VPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEVM
EHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHDF
FNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQV
VATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGQQFL
INHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDWA
DEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARDK
NHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
KSAAFLLQKRWTGMNFGEKPQQGGKQ"
misc_feature 1088..1090
/label=N358Q glycosylation mutation
/note="N358Q glycosylation mutation"
CDS 1829..2539
/codon_start=1
/gene="mgfp5"
/product="GFP with folding enhancement mutations"
/label=mgfp5
/note="suitable for expression in plants due to removal of
a cryptic intron"
/translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
FKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
CDS 2546..2563
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 2595..2847
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 2598..2850
/label=nos (nopaline synthase) 3UTR (polyA signal)
/note="nos (nopaline synthase) 3UTR (polyA signal)"
misc_feature 2869..2893
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
misc_feature 2888..2913
/label=right border T-DNA repeat
/note="right border T-DNA repeat"
misc_feature complement(3954..4954)
/label=STA region from pVS1 plasmid
/note="STA region from pVS1 plasmid"
CDS 4193..4822
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 5256..6323
/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(5547..6547)
/direction=LEFT
/note="pVS1-REP; replication origin from pVS1"
rep_origin 6389..6583
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 6927..7067
/label=bom
/note="basis of mobility region from pBR322"
misc_feature complement(6957..7217)
/label=bom site from pBR322
/note="bom site from pBR322"
rep_origin complement(7253..7841)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
rep_origin complement(7357..7637)
/direction=LEFT
/label=pBR322 origin of replication
/note="pBR322 origin of replication"
CDS complement(7928..8722)
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=aminoglycoside phosphotransferase
/note="aadA (kanamycin resistance) gene amplified from
pIG121Hm"
/protein_id="AAF65349.1"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 9147..9172
/label=left border repeat from C58 T-DNA
/note="left border repeat from C58 T-DNA"
misc_feature 9147..9171
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
misc_feature 9239..9447
/label=CaMV 3UTR (polyA signal)
/note="CaMV 3UTR (polyA signal)"
polyA_signal 9249..9423
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(9463..10488)
/codon_start=1
/product="hygromycin phosphotransferase"
/label=hygromycin phosphotransferase
/note="hptII (hygromycin resistance) gene"
/protein_id="AAF65350.1"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
regulatory complement(10524..11331)
/regulatory_class="promoter"
/note="CaMV35S2; CaMV 35S promoter, duplicated"
promoter complement(10556..11233)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 11424..11445
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
regulatory 11452..11506
/regulatory_class="promoter"
/note="PlacZ; lacZ promoter"
promoter 11460..11490
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11498..11514
/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 11522..11538
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 11534..11767
/codon_start=1
/product="LacZ alpha fragment"
/label=LacZ alpha fragment
/protein_id="AAF65348.1"
/translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
misc_feature 11548..11604
/label=MCS
/note="pUC18/19 multiple cloning site"
misc_feature 11576..11626
/note="pUC18 MCS; polylinker"
primer_bind complement(11608..11624)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 11818..12355
/regulatory_class="promoter"
/note="CaMV35S; 35S promoter from CaMV"
promoter 12001..12346
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
