Part:BBa_K4673031
b-lactamase coding region is an ampicillin resistance gene
B-lactamases are enzymes produced by bacteria that offer multi-resistance to beta-lactam antibiotics, including cephamycins, cephalosporins, monobactams, penicillins, and carbapenems (ertapenem). β-lactamase provides antibiotic resistance by breaking the molecular structure of antibiotics containing a common element—a four-atom ring known as a β-lactam ring. The β-lactamase is able to hydrolyze and open the β-lactam ring through hydrolysis, thereby inactivating the antibiotic.
In our team’s project, we designed the β-lactamase coding region on the pGEM-T EASY vector that codes for the ampicillin resistance gene. It was considered as the second screening mechanism that can be used to confirm whether the surviving colonies in the results of blue-white screening took up the plasmid that contains the pGEM-T EASY vector during the transformation. Since the pGEM-T EASY vector contains the β-lactamase coding region, E.coli DH5α colonies with well-transformed vectors should survive under the ampicillin-containing environment. Thus, the β-lactamase coding region is helpful for our team to confirm that the white colonies in the culture medium contain the pGEM-T EASY vector.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 542
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 542
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 542
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 542
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 716
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