Basic Vector Information
pGreenII 0000 (No LUC) vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pGreenII 0000 (No LUC) vector Sequence
LOCUS Exported 3294 bp DNA circular SYN 17-NOV-2023
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pGreenII 0000 (No LUC)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3294)
TITLE Plasmids to assay for nuclease cleavage in planta
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 3294)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
FEATURES Location/Qualifiers
source 1..3294
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 30..54
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 159..176
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 293..314
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 329..359
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 367..383
/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 372..394
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 391..407
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 391..407
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 403..768
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
TDRPSQQLRSLNGEWKL"
primer_bind 426..446
/label=T3
/note="T3 promoter, forward primer"
promoter 428..446
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 459..566
/label=MCS
/note="pBluescript multiple cloning site"
primer_bind 483..499
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 483..499
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(533..549)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(534..550)
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind complement(574..593)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(575..593)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(603..620)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(603..619)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(612..634)
/label=M13/pUC Forward
/note="In lacZ gene"
misc_feature 784..806
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA
(truncated)"
rep_origin 933..1368
/label=pSa ori
/note="origin of replication from bacterial plasmid pSa"
primer_bind complement(1648..1667)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 1659..2474
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(1732..1751)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 2645..3233
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3134..3153
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
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