Vectors

From DictyWiki

This page is a summary of Dicty cloning vectors.

Vector Name
Promoter || Fusion || Drug Resistance || Extrachr / Integrative (ORI) || PMID || GenBank Acc

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Contents 1 pA15DBamGAL 2 pA15GFP 3 pA15LUC 4 p1ABsr8 5 pATANB43 6 pB10TP1 7 pB10TP2 8 pB15-GFP 9 pB17S 10 pBIG 11 pBORB 12 pDdGal15 13 pDdGal16 14 pDdGal17 15 pDE104 16 pDE109 17 pDEX H 18 pEZTN:tetR-bsR 19 pDIV1 20 pDIV2 21 pDJS 22 pDV-NTAP 23 pDV-CGFP 24 pDNeoII 25 pDXA-3C 26 pDXA-3D 27 pDXA-3H 28 pDXA-FLAG 29 pDXA-GFP 30 pDXA-GFP2 31 pDXA-HC 32 pDXA-HY 33 pDXD-3C 34 pDXD-3H 35 pEXP4(+) 36 pEXP5(+) 37 pfeI 38 pfeII 39 pJK1 40 pLittle 41 pLD1A15SN 42 pMB74 43 pMUW161 44 pnDEL 45 pPAV 46 pPT130 47 pREP 48 pTIKL 49 pTS1 50 pTX-FLAG 51 pTX-GFP 52 pV18-Tn5 53 pVEII 54 pYU20

pA15DBamGAL

act15^prom || gene - lacZ || neoR || Integrative (no Dicty ORI?) || PMID 2198542

• Compared to pDdlac-1 (Dingermann et al 1989), removes fragment of that caused instability of the plasmid
  
• Removes most of the act15 promoter, keeping the ATG and other basal expression elements

pA15GFP

act15^prom ||gene - GFP || neoR || Integrative || PMID 7489901 || GenBankAcc

• C-terminal GFP fusion

pA15LUC

Promoter: None || Luciferase(act15P/act15T) || neoR || Integrative? (no ORI?) || Lloyd et al 1990 Dev Genet 11:391 || GenBankAcc

p1ABsr8

act15^prom || gene - GFP || bsR || Extrachr (Ddp2 ORI) || PMID 10769206 || GenBank Acc

• Derived from pB15-GFP where the SacI site has been inactivated (pB15GFPXSac)
• neoR has been replaced with bsR
• C-terminal GFP fusion vector

pATANB43

Promoter: None || Fusion: None || neoR (act6P) || Ddp1 || PMID 2813371 || GenBank Acc

pB10TP1

Promoter || Fusion || neoR || Extrachr / Integrative (ORI) || PMID 3436528 || GenBank Acc

pB10TP2

Promoter || Fusion || neoR || Extrachr / Integrative (ORI) || PMID 3436528 || GenBank Acc

pB15-GFP

act15^prom || gene - GFP || neoR || Extrachr (Ddp2 ORI) || PMID 10769206 || GenBank Acc

• Derived from pDXA-3C
• C-terminal GFP fusion vector

pB17S

act15 || gene-HA || neoR || Extrachr / Integrative (ORI) || PMID 15851518 || GenBank Acc

• Not original reference. Authors cite PMID 7557400 but this does not mention pB17S.

pBIG

act15 promoter || Fusion: None || neoR (act6P) || Extrachr (Ddp1 ORI) || PMID 7691416 || GenBank Acc
Notes:

• vector sequence from pBluescript II SK

pBORB

act15 promoter || Fusion: None || neoR (act6P) || Integrative || PMID ? || GenBank Acc
Notes:

• created from memory in Rex's lab, came across in PMID 8816999 where it's not much described except neoR.

pDdGal15

act15^prom || gene - lacZ || neoR || Integrative (no Dicty ORI?) || PMID 2198542

• Compared to pDdlac-1 (Dingermann et al 1989), removes fragment of that caused instability of the plasmid

pDdGal16

act15^prom || gene - lacZ || neoR || Integrative (no Dicty ORI?) || PMID 2198542

• Compared to pDdlac-1, (Dingermann et al 1989) removes fragment of that caused instability of the plasmid

pDdGal17

act15^prom || gene - lacZ || neoR || Integrative (no Dicty ORI?) || PMID 2198542

• Compared to pDdlac-1, (Dingermann et al 1989) removes fragment of that caused instability of the plasmid

pDE104

Integrative vector with hygromycin selection
Promoter || Fusion || Hygromycin (act15P/act15T) || Integrative (no Dicty ORI) || PMID 25460565 || GenBank Acc
Notes: Hygromycin selection does always not works reproducibly

pDE109

Extrachromosomal vector with hygromycin selection
Promoter || Fusion || Hygromycin (act15P/act15T) || Extrachr (Ddp2 ORI) || PMID 25460565 || GenBank Acc

pDEX H

[act15] || GFP || [act6]:neoR || Extrachr / Integrative (ORI) || PMID 1328260 || GenBank Acc
Notes: based on pIC20R; from PMID 7954798 I belive the pDEX plasmids are integrative (PF)

pEZTN:tetR-bsR

Promoter || Fusion || [act15]:BSR || Integrative (ORI) || PMID 12954783 || GenBank Acc Notes: Generates a transposable blasticidin resistance (bsr) cassette

pDIV1

3.8 kb spanning the pyr5-6 gene cloned into pGEM3 (Promega)|| Integrative (ORI) || PMID 1326764|| GenBank Acc
Notes: integrative plasmid with the pyr5-6 selection marker that selects for ura+ prototrophy

pDIV2

3.8 kb spanning the pyr5-6 gene cloned into pGEM3 (Promega)|| Integrative (ORI) || PMID 1326764|| GenBank Acc
Notes: integrative plasmid with the pyr5-6 selection marker that selects for ura+ prototrophy - the pyr5-6 gene is in opposite direction as compared to pDIV1

pDJS

[dscB](discoidin) || Fusion || neoR || Extrachr / Integrative (ORI) || PMID 15289548 (not the primary reference) || GenBank Acc

pDV-NTAP

[act15] || gene -GFP || neoR || Extrachr || PMID 17296313 || GenBank [1] EF028665 Notes: Part of a series of TAP-tagged +/- GFP vectors (see pDV series page)

pDV-CGFP

[act15] || gene -GFP || neoR || Extrachr || PMID 17296313 || GenBank [2] EF028667 Notes: Part of a series of TAP-tagged +/- GFP vectors (see pDV series page)

pDNeoII

act6^prom || no fusion || neoR || Integrative || PMID 2832153

pDXA-3C

act15^prom || gene - myc || neoR || Extrachr (Ddp2 ORI) || PMID 7557400 || GenBank X85118

pDXA-3D

act15 || gene - myc || neoR || Extrachr (Ddp2 ORI) || PMID 14551196 || GenBank
Notes: Altered pDXA-3C; ATG upstream of the cDNA insertion site removed

pDXA-3H

act15^prom || gene - 8xHis || neoR || Extrachr (Ddp2 ORI) || PMID 7557400 || GenBank X85119

pDXA-FLAG

act15^prom || FLAG - gene - myc || neoR || Extrachr? (Ddp2 ORI) || PMID 11078649 Notes: C terminal myc tag not translated if the cDNA provides its own stop codon.

pDXA-GFP

act15^prom || 7xHis-GFP fusion - myc || neoR || Extrachr (Ddp2 ORI) || PMID 11078649
Notes:

• GFP fusion is at C terminus if KpnI is used; at N terminus if SacI, BamHI, XhoI are used.
  
• 8xHis tag always N terminal.
  
• GFP's C terminal myc tag is not recogized by 9E10 antibody.

pDXA-GFP2

act15^prom || 8xHis - GFP fusion- myc || neoR || Extrachr (Ddp2 ORI) || PMID 11078649 Notes:

• GFP fusion is at C terminus if KpnI is used; at N terminus if SacI, BamHI, XhoI are used.
  
• 8xHis tag always N terminal.
  
• GFP's C terminal myc tag is not recogized by 9E10 antibody.

pDXA-HC

act15^prom || 7xHis - gene - myc || neoR || Extrachr (Ddp2 ORI) || PMID 7557400 || GenBank X85123 Notes: C terminal myc tag not translated if the cDNA provides its own stop codon.

pDXA-HY

act15^prom || 8xHis - gene - YL1/2 || neoR || Extrachr (Ddp2 ORI) || PMID 7557400 || GenBank X85122

pDXD-3C

dscB^prom || gene - myc || neoR || Extrachr (Ddp2 ORI) || PMID 7557400 || GenBank X85121

pDXD-3H

dscB^prom || gene - 8xHis || neoR || Extrachr (Ddp2 ORI) || PMID 7557400 || GenBank X85120

pEXP4(+)

act15 || Fusion || neoR || Extrachr (Ddp1 ORI) || PMID 2813371, PMID 7926779 || GenBank EF028663

Notes:

• Based on pAT153 (see PMID 17296313)
• Then on pATSP, which is extrachromosomal, mentioned in PMID 2813371.
• pATPS leads via pEXP1, 2, 3 to EXP4(+) (PMID 7926779)
• Other references: PMID 16079174, PMID 12686622

pEXP5(+)

act15 || Fusion || neoR || Extrachr (Ddp1 ORI) || PMID 17296313 || GenBank EF028664

Notes:

• Based on pEXP4(+) (see PMID 17296313)

pfeI

Promoter: None || Fusion: None || Bleomycin (phleomycin)(act15P/act8T) || Extrachr (Ddp2 ORI) || PMID 1953786 || GenBank Acc

pfeII

Promoter: None || Fusion: None || Bleomycin (phleomycin)(act15P/act8T) || Extrachr (Ddp2 ORI) || PMID 1953786 || GenBank Acc

pJK1

Promoter: act15; 2H3 terminator || Fusion: None || neoR (act6P) || Ddp1 || PMID 1348970 || GenBank Acc
Notes

• Based on pATANB43

pLittle

act15 promoter || Fusion: None || neoR (act6P) || Extrachr (Ddp1 ORI) || PMID 8567716 || GenBank Acc
Notes:

• derivative of pBIG
• differs from pBIG in that a 1.5-kb KpnI fragment in the Ddpl portion of pBIG was removed.

pLD1A15SN

act15 promoter || Fusion: None || neoR (act6P) || Extrachr (Ddp2 ORI) || PMID: 10953006 || GenBank Acc
Notes:

• expression plasmid used for suppressor screen

pMB74

act15 promoter || Fusion: might have GFP (MB74-GFP) || neoR, || Extrachr (ORI?) || PMID 15827084, 17040207|| GenBank Acc
Notes:

• has been modified to have hygR instead of neoR (PMID 17040207)
• based on MB12NEO PMID 10094924

pMUW161

Bleomycin casette with the act6 promoter and the act8 terminator
Promoter: None || Fusion: None || Bleomycin (act6P/act8T) || Extrachr (Ddp2 ORI) || PMID 1720754 || GenBank Acc

pnDEL

act15 promoter || Fusion: None || neoR (act6P)? || Integrative || PMID 3244804 || GenBank Acc

pPAV

act15 promoter || Fusion: CAT:act15T || Drug resistance: none (must be co-transformed) || Extrachr (Ddp2 ORI) || PMID 2825142 || GenBank Acc

pPT130

act15^prom || no fusion || neoR || Extrachr / Integrative (ORI) || PMID 16765443
Notes: No Kozak or ATG.

pREP

Plasmid that confers the Ddp2 ORI trans replication functions || PMID 11078649 (This is not the primary reference)

pTIKL

act15 promoter || Fusion: None || neoR (act6P) || Extrachr (Ddp1 ORI) || PMID 12135375 ||

pTS1

Promoter: None || Fusion: None || neoR/Thymidylate Synthase+ || Extrachr / Integrative (ORI) || PMID 2734099 || GenBank Acc

pTX-FLAG

act15^prom || FLAG - epitope || neoR || Extrachr (Ddp1 ORI) || PMID 11078649
Notes:

• If the cDNA provides its own stop codon the epitope will not be present in the expressed protein.
  
• Ddp1 ORI confers more uniform expression across individual cells.

pTX-GFP

act15^prom || 8xHis - GFP fusion - myc || neoR || Extrachr, (Ddp1 ORI) || PMID 11078649
Notes:

• GFP fusion is at C terminus if KpnI is used; at N terminus if SacI, BamHI, XhoI are used.
  
• 8xHis tag always N terminal.
  
• GFP's C terminal myc tag is not recogized by 9E10 antibody.
  
• Ddp1 ORI confers more uniform expression across individual cells.

pV18-Tn5

Can be used to transfom non-axenic strains
Promoter || Fusion || noeR(V18P/P8A7T) || Extrachr / Integrative (ORI) || PMID 9007012 || GenBank Acc

pVEII

For repressible expression; folate-inducible
dscB^prom (discoidin gamma) || no fusion || neoR || Integrative (ORI) || PMID 1475184 || GenBank Acc

pYU20

act15^prom neoR || Extrachr (ORI ?) || PMID 17664333

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