<jupyter_start><jupyter_text>filipa3_2018-01-23 (pMEC1137)This vector expresses four genes and was assembled from four single gene expression cassettes:Gene | Enzyme | Acronym | Cassette-------------------------------------------------- |-------------------|---|-----|[ScTKL1](http://www.yeastgenome.org/locus/S000006278/overview) |Transketolase |TKL | [pYPK0_PDC1_ScTKL1_RPS19b](pYPK0_PDC1_ScTKL1_RPS19b.ipynb)[ScRPE1](http://www.yeastgenome.org/locus/S000003657/overview) |xylitol dehydrogenase |RPE | [pYPK0_RPS19b_ScRPE1_RPS19a](pYPK0_RPS19b_ScRPE1_RPS19a.ipynb)[ScRKI1](http://www.yeastgenome.org/locus/S000005621/overview) |Xylulokinase |RKI | [pYPK0_RPS19a_ScRKI1_TPI1](pYPK0_RPS19a_ScRKI1_TPI1.ipynb)[CiGXF1](http://mic.sgmjournals.org/content/154/6/1646.full) |Transaldolase |GXF | [pYPK0_TPI1_CiGXF1_ENO2](pYPK0_TPI1_CiGXF1_ENO2.ipynb) [Yeast Pathway Kit Standard Primers](ypk_std_primers.ipynb)     PDC1_ScTKL1_RPS19btp                RPS19b_ScRPE1_RPS19atp                              RPS19a_ScRKI1_TPI1tp                                            TPI1_CiGXF1_ENO2tp                                                            pYPK0-PDC1-ScTKL1-RPS19b-ScRPE1-RPS19a-ScRKI1-TPI1-CiGXF1-ScENO2<jupyter_code>from pydna.all import *
p567,p577,p468,p467,p568,p578,p775,p778,p167,p166 = parse("yeast_pahtway_kit_standard_primers.txt")
pYPKp3 = read("pYPKp3.gb")
from Bio.Restriction import EcoRV
p1 =read("pYPK0_PDC1_ScTKL1_RPS19b.gb")
p2 =read("pYPK0_RPS19b_ScRPE1_RPS19a.gb")
p3 =read("pYPK0_RPS19a_ScRKI1_TPI1.gb")
p4 =read("pYPK0_TPI1_CiGXF1_ENO2.gb")
pcr( p775, p778, p3)
cas1  =pcr( p577, p778, p1)
cas2  =pcr( p775, p778, p2)
cas3  =pcr( p775, p778, p3)
cas4  =pcr( p775, p578, p4)
lin_vector = pYPKp3.linearize(EcoRV)
asm = Assembly( [lin_vector, cas1, cas2, cas3, cas4], limit = 61) # This limit is important!
asm
candidate = asm.assemble_circular()[0]
candidate.figure()
asm.assemble_circular()[0].cseguid()<jupyter_output><empty_output>
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