Plasmids

Plasmids

Overview of plasmids

The following overview lists the plasmids used in the course of this work together with a description of their generation and of essential features. All inserts are characterized by the positions of their first and last nucleotides which are flanked by the (introduced) restriction sites. In the case of deletion mutants the positions of the deleted nucleotides are also given. All positions are given with respect to the first (+1 ) nucleotide of the respective open reading frame. Table 4.2. Overview of plasmids.

Plasmid name (lab-

designation)

Description of the plasmid [source or reference]

pUS-Mev

6304 bp vector derived from pBluescript SK- _{(Stratagene, La Jolla, USA) by cloning 1400 bp} of H. salinarum sequence containing the bop gene between BamHI and HindIII sites and 1994 bp of H. volcanii sequence containing the mevinolin resistance gene between HindIII and

XhoI sites of the multiple cloning site (MCS) [Schweiger, 1996; Pfeiffer et al., 1999] pMKK101

(pM@bop1)

6689 bp vector derived from pUS-Mev by replacing the bop-containing sequence between

BamHI and HindIII sites with the BamHI/HindIII-digested bop-locus insert: -387 to +1390 (bop: 789 bp) [this work]

pMKK012 (pM@∆htr12)

5557 bp vector derived from pUS-Mev by replacing the bop-locus sequence between BamHI and HindIII sites with the BamHI/HindIII-digested ∆htr12 insert: -292 to +1620 but lacking +1 to +1266 (htr12: 1263 bp) [this work]

pMKK112

(pM@β∆htr12) between the 7819 bp vector derived from pMKK012 by insertion of the SpeI and SacI sites: -141 to +2145 (bgaH: 1992 bp) [this work]SpeI/SacI-digested bgaH-locus pMKK103

(pM@β∆htr3)

7713 bp vector derived from pMKK112 by replacing the ∆htr12 insert between BamHI and

HindIII sites with the BamHI/HindIII-digested ∆htr3 insert: -297 to +2669 but lacking -5 to +2421 (htr3: 2418 bp) [this work]

pMKK104 (pM@β∆htr4)

7756 bp vector derived from pMKK112 by replacing the ∆htr12 insert between SpeI and

HindIII sites with the SpeI/HindIII-digested ∆htr4 insert: -302 to +2627 but lacking -3 to +2337 (htr4: 2337 bp) [this work]

pMKK105 (pM@β∆htr5)

7759 bp vector derived from pMKK112 by replacing the ∆htr12 insert between SpeI and

HindIII sites with the SpeI/HindIII-digested ∆htr5 insert: -300 to +2732 but lacking -3 to +2437 (htr5: 2433 bp) [this work]

pMKK106 (pM@β∆htr6)

7776 bp vector derived from pMKK112 by replacing the ∆htr12 insert between BamHI and

HindIII sites with the BamHI/HindIII-digested ∆htr6 insert: -301 to +2674 but lacking -1 to +2371 (htr6: 2370 bp) [this work]

pMKK107 (pM@β∆htr7)

7772 bp vector derived from pMKK112 by replacing the ∆htr12 insert between BamHI and

HindIII sites with the BamHI/HindIII-digested ∆htr7 insert: -300 to +1939 but lacking +1 to +1641 (htr7: 1638 bp) [this work]

pMKK108 (pM@β∆htr8)

7719 bp vector derived from pUS-Mev by replacing the bop-locus sequence between BamHI and HindIII sites with the BamHI/HindIII-digested ∆htr8 insert: -315 to +2163 lacking +1 to +1932 (htr8: 1932 bp). Additionally the bgaH-locus sequence was inserted between the SpeI and SacI sites as in the construction of pMKK112 [this work]

pMKK109 (pM@β∆htr9)

7830 bp vector derived from pUS-Mev by replacing the bop-locus sequence between BamHI and HindIII sites with the BamHI/HindIII-digested ∆htr9 insert: -328 to +1774 but lacking +3 to +1449 (htr9: 1446 bp). Additionally the bgaH-locus sequence was inserted between the

Plasmid name (lab-

designation)

Description of the plasmid [source or reference]

pMKK110 (pM@β∆htr10)

7935 bp vector derived from pUS-Mev by replacing the bop-locus sequence between BamHI and HindIII sites with the BamHI/HindIII-digested ∆htr10 insert: -402 to +1830 but lacking +1 to +1470 (htr10: 1470 bp). Additionally the bgaH-locus sequence was inserted between the SpeI and SacI sites as in the construction of pMKK112 [this work]

pMKK111 (pM@β∆htr11)

7817 bp vector derived from pMKK112 by replacing the ∆htr12 insert between SpeI and

HindIII sites with the SpeI/HindIII-digested ∆htr11 insert: -345 to +1664 but lacking -3 to +1356 (htr11: 1359 bp) [this work]

pMKK113 (pM@β∆htr13)

7777 bp vector derived from pMKK112 by replacing the ∆htr12 insert between BamHI and

HindIII sites with the BamHI/HindIII-digested ∆htr13 insert: -296 to +1576 but lacking +4 to +1273 (htr13: 1272 bp) [this work]

pMKK100 (pM@βMaster)

7213 bp vector derived from pUS-Mev by insertion of the SpeI/SacI-digested bgaH-locus between the SpeI and SacI sites: -141 to +2145 (bgaH: 1992 bp). Furthermore the bop- containing sequence between BamHI and XbaI sites was replaced with the BamHI/XbaI- digested MCS-1 insert containing a multiple cloning site (MCS) especially designed for this vector to contain a maximum number of unique cutting sites for the most common restriction enzymes (48 bp between BamHI and XbaI sites) [this work]

pMKK114 (pM@β∆htr14)

7877 bp vector derived from pMKK100 by replacing the MCS sequence between BamHI and

HindIII sites with the BamHI/HindIII-digested ∆htr14 insert: -384 to +2196 but lacking +1 to +1876 (htr14: 1884 bp) [this work]

pMKK115 (pM@β∆htr15)

7930 bp vector derived from pUS-Mev by replacing the bop-locus sequence between BamHI and HindIII sites with the BamHI/HindIII-digested ∆htr15 insert: -326 to +2338 but lacking +5 to +1911 (htr15: 1911 bp). Additionally the bgaH-locus sequence was inserted between the SpeI and SacI sites as in the construction of pMKK112 [this work]

pMKK116 (pM@β∆htr16)

7762 bp vector derived from pUS-Mev by replacing the bop-locus sequence between SpeI and

HindIII sites with the SpeI/HindIII-digested ∆htr16 insert: -324 to +2649 but lacking +2 to +2379 (htr16: 2379 bp). Additionally the bgaH-locus sequence was inserted between the SpeI and SacI sites as in the construction of pMKK112 [this work]

pMKK117 (pM@β∆htr17)

7755 bp vector derived from pMKK112 by replacing the ∆htr12 insert between BamHI and

HindIII sites with the BamHI/HindIII-digested ∆htr17 insert: -295 to +1908 but lacking +1 to +1622 (htr17: 1611 bp) [this work]

pMKK118 (pM@β∆htr18)

7962 bp vector derived from pUS-Mev by replacing the bop-locus sequence between SpeI and

HindIII sites with the SpeI/HindIII-digested ∆htr18 insert: -459 to +2788 but lacking -1 to +2451 (htr18: 2451 bp). Additionally the bgaH-locus sequence was inserted between the SpeI

and SacI sites as in the construction of pMKK112 [this work] pMKK119

(pM@βhtr14)

9757 bp vector derived from pMKK100 by replacing the MCS sequence between BamHI and

HindIII sites with the BamHI/HindIII-digested htr14-locus sequence: -384 to +2196 (htr14: 1884 bp) [this work]

pMKK121 (pM@ β∆oe1534)

7798 bp vector derived from pMKK100 by replacing the MCS sequence between BamHI and

HindIII sites with the BamHI/HindIII-digested ∆oe1534 insert: -299 to +1093 but lacking +1 to +771 (oe1534: 771 bp) [this work]

pMKK102 (pM@βhop2)

9448 bp vector derived from pMKK100 by replacing the MCS sequence between BamHI and

HindIII sites with the BamHI/HindIII-digested hop-locus sequence: -577 to +1698 (hop: 825 bp) [this work]