PM-YC3.6-LTI6b vector (V004859)

Basic Vector Information

Vector Name:
PM-YC3.6-LTI6b
Antibiotic Resistance:
Streptomycin
Length:
13001 bp
Type:
Binary expression vector
Replication origin:
ori
Source/Author:
Krebs M, Held K, Binder A, Hashimoto K, Den Herder G, Parniske M, Kudla J, Schumacher K.
Promoter:
MAS

PM-YC3.6-LTI6b vector Vector Map

PM-YC3.6-LTI6b13001 bp6001200180024003000360042004800540060006600720078008400900096001020010800114001200012600RB T-DNA repeatCAP binding sitelac promoterlac operatorM13 revAtUBQ10CFPCBPVN173Lti6bE9 terminatorM13 fwdMAS terminatorNeoR/KanRMAS promoterLB T-DNA repeatSmRoribompVS1 oriVpVS1 RepApVS1 StaA

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 ℃.

PM-YC3.6-LTI6b vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_29586       13001 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Binary expression vector PM-YC3.6-LTI6b, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13001)
  AUTHORS   Krebs M, Held K, Binder A, Hashimoto K, Den Herder G, Parniske M, 
            Kudla J, Schumacher K.
  TITLE     FRET-based genetically encoded sensors allow high-resolution live 
            cell imaging of Ca(2) dynamics
  JOURNAL   Plant J. 69 (1), 181-192 (2012)
  PUBMED    21910770
REFERENCE   2  (bases 1 to 13001)
  AUTHORS   Krebs M, Held K, Binder A, Hashimoto K, Den Herder G, Parniske M, 
            Kudla J, Schumacher K.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-OCT-2011) Department of Developmental Biology, Centre 
            for Organismal Studies Heidelberg, Im Neuenheimer Feld 230, 
            Heidelberg 69120, Germany
REFERENCE   3  (bases 1 to 13001)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 13001)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant J."; 
            date: "2012"; volume: "69"; issue: "1"; pages: "181-192"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-OCT-2011) Department of Developmental Biology, Centre for 
            Organismal Studies Heidelberg, Im Neuenheimer Feld 230, Heidelberg 
            69120, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..13001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(114..138)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    303..324
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        339..369
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    377..393
                     /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     401..417
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      444..1089
                     /label=AtUBQ10
                     /note="AtUBQ10"
                     /regulatory_class="promoter"
     CDS             1108..1791
                     /label=CFP
                     /note="cyan variant of GFP"
     CDS             2248..2325
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             2548..3066
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     misc_feature    3124..3288
                     /label=Lti6b
                     /note="Lti6b"
     terminator      3304..3945
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     primer_bind     complement(3958..3974)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(4556..4808)
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     CDS             complement(5007..5798)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(5828..6208)
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     misc_feature    complement(6580..6604)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             7132..7920
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     rep_origin      8167..8755
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(8941..9081)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(9425..9619)
                     /direction=LEFT
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9688..10752)
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(11189..11815)
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"

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