Table 2.
3×FLAG translational-mapping plasmids
| Plasmid | Vector | Inserta | Restriction sites | Methodb |
|---|---|---|---|---|
| pPOP/3×FLAG | pHW2000/PR8/PB1 | pOOO/3×FLAG (nt 71–444) | BstAPI, BfuAI | P |
| pMPM/3×FLAG | pDP2002/Mx09/PB1 | pDP2002/Mx09/PB1 (nt 1–391); pPPP/3×FLAG (nt 392–445) | BsaAI, BstAPI | O |
| pOPO/3×FLAG | pHW2000/OH07/PB1 | pHW2000/OH07/PB1 (nt 1–391); pPPP/3×FLAG (nt 392–445) | BsaAI, BstAPI | O |
| pPPM/3×FLAG | pDP2002/Mx09/PB1 | pPPP/3×FLAG (nt 1–391); pDP2002/Mx09/PB1 (nt 392–445) | BsaAI, BstAPI | O |
| pPOM/3×FLAG | pDP2002/Mx09/PB1 | pPOP/3×FLAG (nt 1–391); pDP2002/Mx09/PB1 (nt 392–445) | BsaAI, BstAPI | O |
| pPPO/3×FLAG | pHW2000/OH07/PB1 | pPPO/3×FLAG (nt 1–391); pH 2002/OH07/PB1 (nt 392–445) | BsaAI, BstAPI | O |
| pPOO/3×FLAG | pHW2000/OH07/PB1 | pPOO/3×FLAG (nt 1–391); pHW2000/OH07/PB1 (nt 392–445) | BsaAI, BstAPI | O |
| pMOO/3×FLAG | pHW2000/OH07/PB1 | pMOM/3×FLAG (nt 1–445) | BsaAI, BstAPI | P |
| pOOM/3×FLAG | pDP2002/Mx09/PB1 | pOOO/3×FLAG (nt 1–445) | BsaAI, BstAPI | P |
| pMPO/3×FLAG | pHW2000/OH07/PB1 | pMPM/3×FLAG (nt 1–445) | BsaAI, BstAPI | P |
| pOPM/3×FLAG | pDP2002/Mx09/PB1 | pOPO/3×FLAG (nt 1–445) | BsaAI, BstAPI | P |
| pMOP/3×FLAG | pHW2000/PR8/PB1 | pMOM/3×FLAG (nt 1–444) | NheI, BfuAI | P |
| pMPP/3×FLAG | pHW2000/PR8/PB1 | pMPM/3×FLAG (nt 1–444) | NheI, BfuAI | P |
| pOOP/3×FLAG | pHW2000/PR8/PB1 | pOOO/3×FLAG (nt 1–444) | NheI, BfuAI | P |
| pOPP/3×FLAG | pHW2000/PR8/PB1 | pOPO/3×FLAG (nt 1–444) | NheI, BfuAI | P |
| pMPM(P1121-2298)/3×FLAG | pMPP/3×FLAG | pMPM/3×FLAG (nt 1–1140) | NheI | S |
| pMPM(P392-1140 M)3×FLAG | pMPM/3×FLAG | pMPP/3×FLAG (nt 1–1140) | NheI | S |
| pMPM(P392-816)/3×FLAG | pDP2002/Mx09/PB1 | pMPM(P392–1140 M)3×FLAG (nt 1–817) | BsaAI, HindIII | P |
| pMPM(P392-586)/3×FLAG | pMPM(P392-816)/3×FLAG | pDP2002/Mx09/PB1 (nt 583–817) | BsmBI, HindIII | P |
| pMPM(P582-816)/3×FLAG | pMPM(P392-816)/3×FLAG | pMPM/3×FLAG (nt 1–582) | BsaAI, BsmBI | P |
| pMO(P267-391)M/3×FLAG | pDP2002/Mx09/PB1 | pMOM/3×FLAG (nt 1–266); pMPM/3×FLAG (nt 267–445) | BsaAI, BstAPI | O |
| pM(P119-252)OM/3×FLAG | pDP2002/Mx09/PB1 | pMPM/3×FLAG (nt 1–252); pMOM/3×FLAG (nt 253–445) | BsaAI, BstAPI | O |
| pMO(P344-816)M/3×FLAG | pDP2002/Mx09/PB1 | pMOM/3 × FLAG (nt 1–336); pMPM(P582-816)/3×FLAG (nt 337–817) | BsaAI, EcoRI, HindIII | P |
| pMO(P267-335)O(P392-816)M/3×FLAG | pDP2002/Mx09/PB1 | pMO(P267-391)M/3×FLAG (nt 1–336); pMOP/3×FLAG (nt 337–817) | BsaAI, EcoRI, HindIII | P |
| pMOMF2UP/3×FLAG | pMPM(P392-816)/3×FLAG | pMO(P267-335)O(P392-816)M/3×FLAG (nt 1–336); pMOM/3×FLAG (nt 337–582) | BsaAI, EcoRI, BsmBI | P |
| pPP(O267-343)P/3×FLAG | pHW2000/PR8/PB1 | pPPP/3×FLAG (nt 71–266); pMO(P344-816)M/3×FLAG (nt 267–444) | BstAPI, BfuAI | O |
| pPR8F2dwn/3×FLAG | pPP(O267-343)P/3×FLAG | pDP2002/Mx09/PB1 (nt 583–817); pHW2000/PR8/PB1 (nt 818–1134) | BsmBI, HindIII, PstI | P |
a
The numbers correspond to the PB1 nucleotides inserted from the parent plasmid(s) into the vector and are in reference to the beginning of the influenza virus 5′ untranslated region prior to the PB1 mRNA sequence; they do not include the 3×FLAG sequence.
b
Method used to generate the insert. P, PCR; O, overlap PCR; S, subcloning.