Basic Vector Information
- Vector Name:
- pGWB765
- Antibiotic Resistance:
- Streptomycin
- Length:
- 12069 bp
- Type:
- Gateway binary vector
- Replication origin:
- ori
- Source/Author:
- Momtaz S, Dutta A, Tanaka Y, Aboulela M, Nishimura K, Sugiura S, Niwa T, Maeo K, Goto-Yamada S, Kimura T, Ishiguro S, Mano S, Nakagawa T.
pGWB765 vector Map
pGWB765 vector Sequence
LOCUS 40924_23889 12069 bp DNA circular SYN 18-DEC-2018
DEFINITION Gateway binary vector pGWB765 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12069)
AUTHORS Momtaz S, Dutta A, Tanaka Y, Aboulela M, Nishimura K, Sugiura S,
Niwa T, Maeo K, Goto-Yamada S, Kimura T, Ishiguro S, Mano S,
Nakagawa T.
TITLE Gateway Binary Vectors with Organelle-Targeted Fluorescent Proteins
for Highly Sensitive Reporter Assay in Gene Expression Analysis of
Plants
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 12069)
AUTHORS Momtaz S, Dutta A, Tanaka Y, Aboulela M, Nishimura K, Sugiura S,
Niwa T, Maeo K, Goto-Yamada S, Kimura T, Ishiguro S, Mano S,
Nakagawa T.
TITLE Direct Submission
JOURNAL Submitted (17-OCT-2018) Contact:Tsuyoshi Nakagawa Shimane
University, Interdisciplinary Center for Science Research; 1060
Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE 3 (bases 1 to 12069)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 12069)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(17-OCT-2018) Contact:Tsuyoshi Nakagawa Shimane University,
Interdisciplinary Center for Science Research; 1060 Nishikawatsu,
Matsue, Shimane 690-8504, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT constructed using pPZP221.
FEATURES Location/Qualifiers
source 1..12069
/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 338..462
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 499..529
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 583..1239
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 1584..1886
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(1930..2054)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
CDS 2071..2091
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2092..2808
/codon_start=1
/label=EGFP
/note="enhanced GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
terminator 2833..3085
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3119..3135)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3143..3159
/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(3167..3197)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3212..3233)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
terminator complement(3269..3521)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(3559..4854)
/codon_start=1
/gene="gpt"
/product="UDP-N-acetylglucosamine: dolichol phosphate
N-acetylglucosamine-1-P transferase"
/label=gpt
/protein_id="BBG75662.1"
/translation="MAARKRASSISIPNKPDPSEPNSAPSEQKMTRKTVSASGEEFRLA
PPKLGVIFVISTLLCSLYLYLLCFHYKVDNELKRSILINAGLSLVGFFVTLKMIPVTAR
YVLRRNMFGFDINKRGTPQGDIKVPESLGIVVGIVFLIVAIIFQYFNFTEDSNWLVEYN
AALASICFMILLGFVDDVLDVPWRVKLVLPSFATLPLLMAYAGHTTIVIPKPLVAYIGL
EVLNLGRIYKLYMGLLAVFCTNSINIHAGLNGLEIGQTVVIAAAILIHNVMQIGASVDP
EYHQAHAFSIFLTQPLMATSLAMLAYNWYPSSVFVGDTYTVFAGMTMAVVGILGHFSET
LLIFFLPQVLNLLLSLPQLAGIVKCPRHRLPRYDPATGLLTGTKDGTLVNVYLRLFGPK
SEKSLCIHLLVFQALACAFCFILRHFLAGWYK"
gene complement(3559..4854)
/gene="gpt"
/label=gpt
promoter complement(4906..5085)
/label=NOS promoter
/note="nopaline synthase promoter"
misc_feature complement(5591..5615)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 6136..6924
/codon_start=1
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 7173..7761
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(7947..8087)
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(8431..8625)
/direction=LEFT
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(8694..9764)
/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(10196..10822)
/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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