Lab. Plant Molecular Genetics - Nakagawa & Hachiya Lab.
Lab. Plant Molecular Genetics - Nakagawa & Hachiya Lab. HOME > Gateway Binary Vector (pGWB)
 
Information of Gateway Binary Vector (pGWB)
 
Gateway Binary Vector with organelle-targeted fluorescent protein >>
Gateway Vector for BiFC >>     Gateway Recycling Cloning >>     Dual-Site Gateway Cloning >>
 
Review (INTECH Open Access): Gateway Vectors for Plant Genetic Engineering:
Overview of Plant Vectors, Application for Bimolecular Fluorescence
Complementation (BiFC) and Multigene Construction
 
attR1-attR2 type

pGWB series (pBI backbone, Kmr and Hygr for bacteria and plant)
pGWB series
 
Complete List of pGWB　PDF download
  • pGWB1-45 (pBI backbone, PNOS:NTPII and P35S:HPT) (List)
  • pGWB203-235 (pBI backbone, PNOS:NTPII and PNOS:HPT) (List)
  • manual Japanese (Word / PDF) manuals　　Word / PDF
  • manual English (Word / PDF) manuals　　Word / PDF
  • Reference    Most Cited Journal of JBB (last five years)

    pGWB1-45 and 203-235 are planed to be deposited at RIKEN in future. We will announce when the deposit is completed.
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    attR1-attR2 type

    ImpGWB series (pPZP backbone, SpcR for bacteria, KmR or HygR or BASTA-R or TunicamycinR for plant)
    ImpGWB series
     
    Complete List of ImpGWB　PDF download
  • pGWB401-461 (KmR, pPZP backbone, PNOS:NTPII) (List)
  • Reference
  • pGWB401-455 are available from Addgene.
  • pGWB401-461 are available from RIKEN BRC Experimental Plant Division.

  • pGWB501-561 (HygR, pPZP backbone, PNOS:HPT) (List)
  • Reference
  • pGWB501-555 are available from Addgene.
  • pGWB501-555 are available from RIKEN BRC Experimental Plant Division.

  • pGWB601-661 (BASTA-R, pPZP backbone, PNOS:bar) (List)
  • Reference
  • pGWB601-661 are available from RIKEN BRC Experimental Plant Division.

  • pGWB701-761 (TunicamycinR, pPZP backbone, PNOS:GPT) (List)
  • Reference
  • Comments on tunicamycin selection (important)

  • manual Japanese (PDF)
  • manual English (PDF)

  • pGWB with organelle-targeted fluorescent protein
  • pGWB with organelle-targeted fluorescent protein RIKEN BRC Experimental Plant Division



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    attR4-attR2 type (Promoter Swapping)

    R4pGWB series (pPZP backbone, SpcR for bacteria, KmR or HygR or BASTA-R or TunicamycinR for plant)
    R4pGWB series
     
    Complete List of R4pGWB　PDF download
  • R4pGWB401-459 (KmR, pPZP backbone, PNOS:NTPII) (List)
  • Reference
  • pGWB401-459 are available from RIKEN BRC Experimental Plant Division.

  • R4pGWB501-559 (HygR, pPZP backbone, PNOS:HPT) (List)
  • Reference
  • pGWB501-559 are available from RIKEN BRC Experimental Plant Division.

  • R4pGWB601-659 (BASTA-R, pPZP backbone, PNOS:bar) (List)
  • Reference
  • pGWB601-659 are available from RIKEN BRC Experimental Plant Division.

  • R4pGWB701-759 (TunicamycinR, pPZP backbone, PNOS:GPT) (List)
  • Reference
  • Comments on tunicamycin selection (important)

  • manual Japanese (PDF)
  • manual English (PDF)

    ● Information concerning pDONR P4-P1R (Donor vector for construction of attL4-promoter-attR1 entry clones)
    The sale of pDONR P4-P1R (MultiSite Gateway Three-Fragment Vector Construction Kit) was terminated. Instead of pDONR P4-P1R, you may use pENTR™ 5'-TOPO™ TA Cloning™ Kit for construction of attL4-promoter-attR1 entry clones.

    https://www.thermofisher.com/order/catalog/product/K59120?ICID=search-k59120


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    attR4-attL1type (Promoter Analysis)

    R4L1pGWB series (pPZP backbone, SpcR for bacteria, KmR or HygR or BASTA-R for plant)
    R4L1pGWB series
     
    Complete List of R4L1pGWB　PDF download
  • R4L1pGWB432-459 (KmR, pPZP backbone, PNOS:NTPII) (List)
  • Reference

  • R4L1pGWB532-559 (HygR, pPZP backbone, PNOS:HPT) (List)
  • Reference

  • R4L1pGWB632-659 (BASTA-R, pPZP backbone, PNOS:bar) (List)
  • Reference
  • MTA for BASTA-R R4L1 vectors Japanese (Word / PDF)
  • MTA for BASTA-R R4L1 vectors English (Word / PDF)

  • R4L1pGWB732-759 (TunicamysinR, pPZP backbone, PNOS:GPT (List)
  • Reference
  • Comments on tunicamycin selection (important)

  • manual Japanese (PDF)
  • manual English (PDF)

  • pGWB with organelle-targeted fluorescent protein
  • pGWB with organelle-targeted fluorescent protein RIKEN BRC Experimental Plant Division

    ● Information concerning pDONR P4-P1R (Donor vector for construction of attL4-promoter-attR1 entry clones)
    The sale of pDONR P4-P1R (MultiSite Gateway Three-Fragment Vector Construction Kit) was terminated. Instead of pDONR P4-P1R, you may use pENTR™ 5'-TOPO™ TA Cloning™ Kit for construction of attL4-promoter-attR1 entry clones.

    https://www.thermofisher.com/order/catalog/product/K59120?ICID=search-k59120


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    Gateway Binary Vector with organelle-targeted fluorescent protein

    pGWB series (attR1-attR2 type, pPZP ackbone, SpcR for bacteria, KmR or HygR or BASTA-R or TunicamycinR for plant)
    R4L1pGWB series (attR4-attL1 type, pPZP backbone, SpcR for bacteria, KmR or HygR or BASTA-R or TunicamycinR for plant)
    Gateway Binary Vector with organelle-targeted fluorescent protein
     
    Complete List of pGWB and R4L1pGWB with organelle-targeted fluorescent protein　PDF download
  • pGWB462-491, R4L1pGWB462-491 (KmR, pPZP backbone, PNOS:NTPII) (List)
  • Reference

  • pGWB562-591, R4L1pGWB562-591 (HygR, pPZP backbone, PNOS:HPT) (List)
  • Reference

  • pGWB662-691, R4L1pGWB662-691 (BASTA-R, pPZP backbone, PNOS:bar) (List)
  • Reference

  • pGWB762-791, R4L1pGWB762-791 (TunicamycinR, pPZP backbone, PNOS:GPT) (List)
  • Reference

  • pGWB with organelle-targeted fluorescent protein RIKEN BRC Experimental Plant Division

  • Comments on tunicamycin selection (important)

    ● Information concerning pDONR P4-P1R (Donor vector for construction of attL4-promoter-attR1 entry clones)
    The sale of pDONR P4-P1R (MultiSite Gateway Three-Fragment Vector Construction Kit) was terminated. Instead of pDONR P4-P1R, you may use pENTR™ 5'-TOPO™ TA Cloning™ Kit for construction of attL4-promoter-attR1 entry clones.

    https://www.thermofisher.com/order/catalog/product/K59120?ICID=search-k59120


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    Gateway Vector for BiFC (not binary)

    Gateway Vector for BiFC (pUC119 backbone, Ampr for bacteria)
    Gateway Vector for BiFC
     
    Complete List of Gateway Vector for BiFC　PDF download
  • pGWnY (nYFP/pUGW2) (List)
  • pnYGW (nYFP/pUGW0)
  • pGWcY (cYFP/pUGW2)
  • pcYGW (cYFP/pUGW0)
  • Reference
  • Reference for former name

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    Gateway Recycling Cloning Ver. 1
    recycle illustration


    Gateway Vector for Recycling Cloning Ver. 1 (pDONR201 backbone, Kmr for bacteria)
    Gateway Recycling Cloning
     
    Complete List of Gateway Vector for Recycling Cloning Ver. 1　PDF download
  • pRED419 (List)
  • pRED418
  • pCON
  • Reference

    NOTE    Multiple use of CaMV35S promoters frequently causes silencing in this system.

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    Gateway Recycling Cloning Ver. 2

    Gateway Vector for Recycling Cloning Ver. 2 (pDONR201 backbone, Kmr for bacteria)
    Gateway Recycling Cloning
     
    Improvements in Ver.2
    (1) Single digestion at RCS prior to LR reaction is enough for Ver.2 (Double digestion at RCS is necessary for Ver. 1).
    (2) Ampicillin selection is available in transformation after LR reaction using pRED419-Amp or pRED418-Amp.
    (3) Chloramphenicol selection is available in transformation after LR reaction using pCON-Cm-rare1 or pCON-Cm-rare2.
    (4) Use of ampicillin (chloramphenicol) greatly enhances selection of correctly constructed clones in transformation.

    Complete List of Gateway Vector for Recycling Cloning Ver. 2　PDF download
  • pRED419-Amp (List)
  • pRED418-Amp
  • pCON-Cm-rare1
  • pCON-Cm-rare2
  • Reference In preparation

    NOTE    Multiple use of CaMV35S promoters frequently causes silencing in this system.

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    R4 Dual-Site (R4DS) Gateway Cloning System

    Cloning of two set of promoter and ORF into a binary vector.
    R4pGWB6xxx-MD8 series (pPZP backbone. SpcR for bacteria. KmR or HygR or BASTA-R or TunicamycinR for plant)
    [Intermediate vector R4pDD6xx-MD8 series (pUC119 backbone. AmpR for bacteria)]
    R4DS illust
     
    Complete List of R4DS Gateway Cloning System　PDF download
  • R4pGWB6401-MD8 to R4pGWB6459-MD8, R4pGWB6400-MD8-n/cYFP (KmR, pPZP backbone, Pos:NPTII) (List)
  • Reference

  • R4pGWB6501-MD8 to R4pGWB6559-MD8, R4pGWB6500-MD8-n/cYFP (HygR, pPZP backbone, Pos:HPT) (List)
  • Reference

  • R4pGWB6601-MD8 to R4pGWB6659-MD8, R4pGWB6600-MD8-n/cYFP (BASTA-R, pPZP backbone, Pos:bar) (List)
  • Reference

  • R4pGWB6701-MD8 to R4pGWB6759-MD8, R4pGWB6700-MD8-n/cYFP (TunicamycinR, pPZP backbone, Pnos:GPT) (List)
  • Reference

  • R4pDD601-MD8 to R4pDD659-MD8, R4pDD600-MD8-n/cYFP (AmpR, pUC119 backbone) (List)
  • Reference

    All vectors of R4DS Gateway Cloning System are available from RIKEN BRC Experimental Plant Division.

    ● Information concerning pDONR P4-P1R (Donor vector for construction of attL4-promoter-attR1 entry clones)
    The sale of pDONR P4-P1R (MultiSite Gateway Three-Fragment Vector Construction Kit) was terminated. Instead of pDONR P4-P1R, you may use pENTR™ 5'-TOPO™ TA Cloning™ Kit for construction of attL4-promoter-attR1 entry clones.

    https://www.thermofisher.com/order/catalog/product/K59120?ICID=search-k59120


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    Dual-Site (DS) Gateway Cloning System (Pnos)

    Cloning of two ORF into binary vector.
    Expression by Pnos (nopaline synthase promoter).
    N-terminal fusion with 14 tags are available.
    N-tarminal and C-terminal fusion with n/cYFP are available.
    pGWB6xxx-MD8-Pnos series (pPZP backbone, SpcR for bacteria, KmR or HygR or BASTA-R or Tunicamycin R for plant)
    [Intermediate vector pDD6xx-MD8-Pnos series (pUC119 backbone. AmpR for bacteria)]
    Pnos system
     
    Complete List of DS Gateway Cloning System (Pnos) PDF download
  • pGWB6402-MD8-Pnos to pGWB6461-MD8-Pnos, pGWB6400-MD8-Pnos-N/CY0/2 (KmR, pPZP backbone, Pnos:NPTII) (List)
  • Reference

  • pGWB6502-MD8-Pnos to pGWB6561-MD8-Pnos, pGWB6500-MD8-Pnos-N/CY0/2 (HygR, pPZP backbone, Pnos:HPT) (List)
  • Reference

  • pGWB6602-MD8-Pnos to pGWB6661-MD8-Pnos, pGWB6600-MD8-Pnos-N/CY0/2 (BASTA-R, pPZP backbone, Pnos:bar) (List)
  • Reference

  • pGWB6702-MD8-Pnos to pGWB6761-MD8-Pnos, pGWB6700-MD8-Pnos-N/CY0/2 (TunicamycinR, pPZP backbone, Pnos:GPT) (List)
  • Reference

  • pDD602-MD8-Pnos to pDD661-MD8-Pnos, pDD600-MD8-Pnos-N/CY0/2 (AmpR, pUC119 backbone) (List)
  • Reference

    All vectors of DS Gateway Cloning System (Pnos) are available from RIKEN BRC Experimental Plant Division.
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    pENTR-NeCo-lacZa System

    For preparation of empty vector (negative control construct) in Gateway cloning
     

    Complete List of pENTR-NeCo-lacZa PDF download
     

     
  • Reference

    All pENTR-NeCo-lacZa vectors are available from RIKEN BRC Experimental Plant Division.
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