<jupyter_start><jupyter_text>pYPK3pYPK3 is a version of [pYPK1](pYPK1.ipynb) that has a CEN6/ARS yeast oriand a HphMX4 marker instead of the bleomycin marker.<jupyter_code>from pydna.all import *
pYPK1 =read("pYPK1.gb")
gb =Genbank("<EMAIL>")
githubuser="BjornFJohansson"
gistid = "c5424b7ebbf553c52053"
x=download_text('https://gist.githubusercontent.com/{user}/{gistid}/raw'.format(user=githubuser, gistid=gistid))
pAG32 =read(x)
pAG32<jupyter_output><empty_output><jupyter_text>[pAG32 sequence](https://gist.github.com/BjornFJohansson/c5424b7ebbf553c52053)<jupyter_code>p150,p149,p325,p324,p678,p666 = parse_primers('''
>150_MX4fwd (25-mer) Primer in the Ashbya gossypi TEF promoter in forward direction. contaminated?
AAAATCTTGCTAGGATACAGTTCTC

>149_MX4rev (25-mer) Primer in the Ashbya gossypi TEF terminator in the reverse direction.
ACAAATGACAAGTTCTTGAAAACAA

>325_Hygfwd (27-mer)
GATGTAGGAGGGCGTGGATATGTCCTG

>324_Hygrev (27-mer)
TGCATCATCGAAATTGCCGTCAACCAA

>678_pYPK0_hygfwd: (77-mer)
ctcacgttaagggattttggtcatgagCACATACGATTTAGGTGACACTATAGAAC

>666_pYPK0_hygrev (70-mer)
catctttgacagcttatcatcgataagctCGACTCACTATAGGGAGACC''')
hygmarker1 =pcr( p678, p324, pAG32 )
hygmarker2 =pcr( p325, p666, pAG32 )
from Bio.Restriction import PvuII
vect = pYPK1.linearize(PvuII)
a=Assembly([hygmarker1, hygmarker2, vect], limit =251)
a
candidate = a.assemble_circular()[0]
candidate.figure()
pYPK3=candidate.synced(pYPK1)
pYPK3.stamp()
pYPK3.locus = "pYPK3"
pYPK3.write("pYPK3.gb")<jupyter_output><empty_output><jupyter_text>[DOWNLOAD](pYPK3.gb)<jupyter_code>from pydna.all import *
reloaded=read("pYPK3.gb")
assert reloaded.cseguid() in reloaded.definition<jupyter_output><empty_output>
StarcoderdataJupyterStructuredCleanDedup