pGWB554 vector (V005761)

Basic Vector Information

Vector Name:
pGWB554
Antibiotic Resistance:
Streptomycin
Length:
12684 bp
Type:
Gateway binary vector
Replication origin:
ori
Host:
Plants
Source/Author:
Nakagawa T, Suzuki T, Murata S, Nakamura S, Hino T, Maeo K, Tabata R, Kawai T, Tanaka K, Niwa Y, Watanabe Y, Nakamura K, Kimura T, Ishiguro S.
Promoter:
CaMV35S(long)

pGWB554 vector Map

pGWB55412684 bp6001200180024003000360042004800540060006600720078008400900096001020010800114001200012600RB T-DNA repeatM13 fwdCaMV 35S promoterattR1lac UV5 promoterCmRccdBattR2mRFP1NOS terminatorM13 revlac operatorlac promoterCAP binding siteNOS terminatorHygRNOS promoterLB T-DNA repeatSmRoribompVS1 oriVpVS1 RepApVS1 StaA

pGWB554 vector Sequence

LOCUS       40924_23314       12684 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway binary vector pGWB554 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12684)
  AUTHORS   Nakagawa T, Suzuki T, Murata S, Nakamura S, Hino T, Maeo K, Tabata 
            R, Kawai T, Tanaka K, Niwa Y, Watanabe Y, Nakamura K, Kimura T, 
            Ishiguro S.
  TITLE     Improved Gateway Binary Vectors: High-Performance Vectors for 
            Creation of Fusion Constructs in Transgenic Analysis of Plants
  JOURNAL   Biosci. Biotechnol. Biochem. (2007) In press
  PUBMED    17690442
REFERENCE   2  (bases 1 to 12684)
  AUTHORS   Nakagawa T, Ishiguro S.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-FEB-2007) Tsuyoshi Nakagawa, Shimane University, 
            Center for Integrated Research in Science; 1060 Nishikawatsu, 
            Matsue, Shimane 690-8504, Japan 
            (E-mail:tnakagaw@life.shimane-u.ac.jp, Tel:81-852-32-6595, 
            Fax:81-852-32-6109)
REFERENCE   3  (bases 1 to 12684)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 12684)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biosci. 
            Biotechnol. Biochem. (2007) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-FEB-2007) Tsuyoshi Nakagawa, Shimane University, Center for 
            Integrated Research in Science"; volume: " 1060 Nishikawatsu, 
            Matsue, Shimane 690-8504, Japan 
            (E-mail:tnakagaw@life.shimane-u.ac.jp, Tel:81-852-32-6595, Fax"; 
            pages: "81-852-32-6109"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     constructed using pPZP221.
FEATURES             Location/Qualifiers
     source          1..12684
                     /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"
     promoter        812..1157
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    1179..1303
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1340..1370
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1424..2080
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2425..2727
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2771..2895)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2906..3580
                     /codon_start=1
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3605..3857
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3891..3907)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3915..3931
                     /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(3939..3969)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3984..4005)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     terminator      complement(4041..4293)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(4447..5469)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGAFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(5521..5700)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    complement(6206..6230)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6751..7539
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      7788..8376
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(8562..8702)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(9046..9240)
                     /direction=LEFT
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9309..10379)
                     /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(10811..11437)
                     /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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