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AMB Express

Table 2 Sequencing results of vector backbones that were subjected to epMEGAWHOP optimization

From: Increasing protein production by directed vector backbone evolution

pET28a(+)-CelA2

pET22b(+)-BSLA

pHY-scarlsberg

bp

Substitution

Region

bp

Substitution

Region

bp

Substitution

Region

 

wt

-

M1

  

wt

-

M1

  

wt

-

M1

 

488

A

-

T

f1 origin

123

T

-

C

T7 term

209

A

-

G

Tet

850

G

-

C

Kan

2007

A

-

G

na

684

A

-

G

Tet

2233

T

-

C

pBR322 ori

3701

A

-

G

Tet

1584

T

-

C

pTet

2396

T

-

C

na

4563

T

-

C

lacI

3238

T

-

C

pAMP

3170

T

-

C

na

4794

C

-

T

lacI

4221

T

-

C

na

5193

A

-

T

T7 prom

     

4332

A

-

-

ori-177

  1. The positions of the mutations in the corresponding vector backbones are based on the sequenced vector backbone prior epMEGAWHOP. Alignments of all three vector systems to the corresponding ‘parents’ are included in Additional file 1: Figure S1. Nucleotide position 1 is the base after the stop codon (ATT, TAA) of the inserted genes.
  2. na: no corresponding function of the gene sequence could be assigned.
  3. Kan/Tet: Kanamycin/Tetracyclin resistance gene.
  4. pTET/pAMP: promoter region Tetracyclin/Ampicilin resistance gene (identified with a promoter prediction tool (Reese 2001)).
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