TABLE 1.
Plasmid clones (not described in Materials and Methods) used in this work
| Plasmid | Distance (bp)
|
Construction | |
|---|---|---|---|
| Between cassettes | From poly(A) site to postcassette | ||
| p<> | 96 | Same as pORgf3·2 (see Materials and Methods) | |
| p<neo> | 588 | 495-bp BglII-RsaI fragment from the neomycin resistance gene of pSV2neo (57) inserted into the BamHI site of p<> | |
| p<ABCD> | 2,098 | 1,330 | As for p<ABCDE> except for a ClaI-SstI fragment inserted into p<> |
| p<ABC> | 1,578 | 806 | BamHI segment of p<ABCDE>inserted into the BamHI site of p<> |
| p<AB> | 941 | 173 | As for p<ABCDE> except for a ClaI-AflIII fragment inserted into p<> (with regeneration of BamHI sites) |
| p<ABC1> | 1,275 | 511 | AflIII-RsaI fragment from p<ABC> inserted into the XbaI site of p<AB> |
| p<ABC2> | 1,260 | 492 | RsaI-XbaI fragment from p<ABC> inserted into the XbaI site of p<AB> |
| p<ABDEF> | 1,781 | 1,013 | ABC segment removed from p<ABCDEF> with BamHI and replaced with the AB BamHI fragment from p<AB> |
| p<DEF> | 866 | SalI fragment from pCBG12.8 (50) inserted into the BamHI site of p<> | |
| p<D> | 605 | SalI-SstI fragment from pCBG12.8 (previously inserted into the BamHI site of p<> in the reverse orientation) re-excised with SmaI and reinserted into the BamHI site of p<> in the native orientation | |
| p<BD> | 843 | 699 | KpnI-BglII PCR fragment extending from −124 to +137 on the map (Fig. 2) joined to a BamHI-KpnI PCR fragment extending from 799 to 1262 on the map by means of compatible sticky-end ligation between the BglII and BamHI sites, with the resulting KpnI fragment blunted into the BamHI site of p<> |
| p<ABD> | 1,453 | 685 | As for p<D> except with the SmaI fragment inserted into the XbaI site of p<AB> |
| pAB<DEF> | 866 | 1,399 | ClaI-AflIII segment of pABCDE inserted into the KpnI site of p<DEF> |
| pAB<neo> | 588 | 1,121 | SmaI fragment of pAB<DEF> replaced with the SmaI fragment from p<neo> |
| p<ABneo> | 1,487 | 719 | SmaI fragment from p<neo> inserted into the XbaI site of p<AB> |
| p<A> | 655 | SmaI fragment of p<> replaced with a SalI fragment from pAB<neo> | |
| pAB<> | 46 | 579 | neo fragment removed from pAB<neo> with SmaI |
| pAB<cat> | 601 | 1,134 | SmaI fragment of pAB<neo> replaced with cat coding sequences (the HindIII-NcoI fragment) from pRSVcat (24) |
| p<cat> | 601 | SalI fragments containing segment AB deleted from pAB<cat> | |
| p<ABcat> | 1,500 | 732 | HindIII-NcoI fragment of pRSVcat inserted into the XbaI site of p<AB> |
| pAP<cat> | 601 | 1,059 | XhoI fragment of pAB<cat> replaced by the SV40 early poly(A) signal (indicated by capital letters) contained in the following oligonucleotide and its complement: 5′-cccggttaACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTggatccccgt (11) |
| pAPΔ<cat> | 594 | 1,052 | As for pAP<cat>, except with the segment coding for the U-rich element (underlined in pAP<cat> deleted |
| pAPrev<cat> | 601 | As for pAP<cat>, with the oligonucleotide inserted in reverse orientation | |
| pAP<neo> | 588 | 1,047 | As for pAP<cat>, with pAB<neo> as the starting construct |
| pA<cat> | 601 | As for pAP<cat>, with no poly(A) signal insertion | |
| p<lac> | 545 | PvuII fragment containing lac and multiple cloning site sequences from pBluescript SKII+ (Stratagene) inserted into the BamHI site of p<> | |
| pAB<lac> | 492 | 1,027 | SmaI fragment of pAB<neo> replaced with the PvuII fragment from pBluescript SKII+ |
| p<ABlac> | 1,393 | 625 | PvuII fragment from pBluescript SKII+ inserted into the XbaI site of p<AB> |
| pD<C2> | 358 | XbaI fragment containing segment D excised from p<D> and inserted into the upstream XhoI site of p<D> to give pD<D>, with the RsaI-XbaI segment of p<ABC> used to replace the SmaI segment of pD<D> | |
| pDAB<C2> | 358 | 862 | ClaI-AflIII segment of pABCDE inserted into the HindIII site of pD<C2> |
| pD<BC2> | 636 | 506 | BC2XhoI-EcoRI fragment of p<ABC2> used to replace the SmaI segment of pD<D> (see pD<C2>) |
| pDA<C2> | 358 | SalI fragment containing segment A (see Materials and Methods) inserted into the HindIII site of pD<C2> | |
| pDA<BC2> | 636 | 506 | As for pDA<C2> except with insertion into pD<BC2> |
| pD<> | 46 | SmaI fragment deleted from pD<D> (see pD<C2>) | |
| pD<A> | 655 | BamHI-SstI fragment from pABCDE inserted into the ApaI site of p<A> | |
| p(AB)sv<cat> | 601 | 1,078 | SalI fragments of pAB<cat>, containing the βH-globin AB region, replaced by the NcoI-BamHI fragment of pRSVcat, containing a comparable region from SV40 |
| p<WXYZ> | 2,389 | 1,906 | Chicken HindIII-BamHI fragment from pCBG15 (63) inserted into the BamHI site of p<> |
| p<W> | 627 | 144 | ApaI-SstI segment of p<WXYZ> inserted at the BamHI site of p<> |
| p<WX> | 1,220 | 737 | SstI-PvuII segment of p<WXYZ> inserted at the XbaI site of p<W> |
| p<WY> | 1,263 | 780 | ApaLI-BglI segment of p<WXYZ> inserted at the XbaI site of p<W> |
| p<WZ> | 1,272 | 789 | BglI-XbaI segment of p<WXYZ> inserted at the XbaI site of p<W> |
| p<WYZ> | 1,927 | 1,444 | ApaLI-XbaI segment of p<WXYZ> inserted at the XbaI site of p<W> |
| pORgf1·2 | 109 | HindIII-XbaI fragment of pORgf3·2 containing the 376-bp G-free cassette replaced with an SstI-ApaI fragment from pRL542 containing a 130-bp G-free cassette | |
| p<neo>1·2 | 588 | SmaI fragment of pORgf1·2 replaced with the SmaI fragment from p<neo> | |
| pAB<neo>1·2 | 588 | 869 | SmaI fragment from p<AB> inserted at the XhoI site of p<neo>1·2 |