Basic Vector Information
- Vector Name:
- pGWB728
- Antibiotic Resistance:
- Streptomycin
- Length:
- 11853 bp
- Type:
- Gateway binary vector
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Tanaka Y, Nakamura S, Kawamukai M, Koizumi N, Nakagawa T.
- Promoter:
- NOS
pGWB728 vector Map
pGWB728 vector Sequence
LOCUS 40924_23789 11853 bp DNA circular SYN 18-DEC-2018
DEFINITION Gateway binary vector pGWB728 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11853)
AUTHORS Tanaka Y, Nakamura S, Kawamukai M, Koizumi N, Nakagawa T.
TITLE Development of a series of gateway binary vectors possessing a
tunicamycin resistance gene as a marker for the transformation of
Arabidopsis thaliana
JOURNAL Biosci. Biotechnol. Biochem. 75 (4), 804-807 (2011)
PUBMED 21512216
REFERENCE 2 (bases 1 to 11853)
AUTHORS Tanaka Y, Nakagawa T.
TITLE Direct Submission
JOURNAL Submitted (12-JAN-2011) Contact:Tsuyoshi Nakagawa Shimane
University, Center for Integrated Research in Science; 1060
Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE 3 (bases 1 to 11853)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11853)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biosci.
Biotechnol. Biochem."; date: "2011"; volume: "75"; issue: "4";
pages: "804-807"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(12-JAN-2011) Contact:Tsuyoshi Nakagawa Shimane University, Center
for Integrated Research in Science; 1060 Nishikawatsu, Matsue,
Shimane 690-8504, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT constructed using pPZP221.
FEATURES Location/Qualifiers
source 1..11853
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(54..78)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 281..297
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 323..447
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 484..514
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 568..1224
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 1569..1871
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(1915..2039)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
CDS 2050..2127
/codon_start=1
/label=CBP
/note="calmodulin-binding peptide"
/translation="KRRWKKNFIAVSAANRFKKISSSGAL"
CDS 2155..2175
/codon_start=1
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
/translation="ENLYFQG"
CDS 2215..2388
/codon_start=1
/label=ProtA
/note="IgG-binding unit of Staphylococcus aureus protein A"
/translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
AEAKKLNDAQAPK"
CDS 2392..2562
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
EAKKLNGAQAPK"
terminator 2617..2869
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(2903..2919)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2927..2943
/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(2951..2981)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2996..3017)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
terminator complement(3053..3305)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(3343..4638)
/codon_start=1
/gene="gpt"
/product="UDP-N-acetylglucosamine: dolichol phosphate
N-acetylglucosamine-1-P transferase"
/label=gpt
/protein_id="BAJ61241.1"
/translation="MAARKRASSISIPNKPDPSEPNSAPSEQKMTRKTVSASGEEFRLA
PPKLGVIFVISTLLCSLYLYLLCFHYKVDNELKRSILINAGLSLVGFFVTLKMIPVTAR
YVLRRNMFGFDINKRGTPQGDIKVPESLGIVVGIVFLIVAIIFQYFNFTEDSNWLVEYN
AALASICFMILLGFVDDVLDVPWRVKLVLPSFATLPLLMAYAGHTTIVIPKPLVAYIGL
EVLNLGRIYKLYMGLLAVFCTNSINIHAGLNGLEIGQTVVIAAAILIHNVMQIGASVDP
EYHQAHAFSIFLTQPLMATSLAMLAYNWYPSSVFVGDTYTVFAGMTMAVVGILGHFSET
LLIFFLPQVLNLLLSLPQLAGIVKCPRHRLPRYDPATGLLTGTKDGTLVNVYLRLFGPK
SEKSLCIHLLVFQALACAFCFILRHFLAGWYK"
gene complement(3343..4638)
/gene="gpt"
/label=gpt
promoter complement(4690..4869)
/label=NOS promoter
/note="nopaline synthase promoter"
misc_feature complement(5375..5399)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 5920..6708
/codon_start=1
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 6957..7545
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(7731..7871)
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(8215..8409)
/direction=LEFT
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(8478..9548)
/codon_start=1
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
CDS complement(9980..10606)
/codon_start=1
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
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