pYPK2 is a version of pYPK1 that has a CEN6/ARS yeast ori and a KanMX4 marker instead of the bleomycin marker.
In [1]:
from pydna.all import *
pYPK1 =read("pYPK1.gb")
gb =Genbank("bjornjobb@gmail.com")
In [2]:
pFA6a_KanMX4 = gb.nucleotide("AJ002680.1")
In [3]:
p478, p666 =parse('''
>478_bleMX4f
aatttaaattataattatttttatagcacgtgatgaaaaggtccccgccgggtc
>666_pYPK0_hygrev (70-mer)
catctttgacagcttatcatcgataagctCGACTCACTATAGGGAGACC''', ds=False)
In [4]:
kanmarker =pcr( p478, p666, pFA6a_KanMX4)
In [5]:
from Bio.Restriction import PvuII
vect = pYPK1.linearize(PvuII)
a=Assembly([kanmarker, vect])
pYPK2 = a.assemble_circular()[0]
pYPK2 = pYPK2.synced(pYPK1)
pYPK2.stamp()
pYPK2.locus = "pYPK2"
pYPK2.write("pYPK2.gb")
In [6]:
from pydna.all import *
reloaded=read("pYPK2.gb")
assert reloaded.cseguid() in reloaded.definition
In [7]: