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EasyClone-MarkerFree: A vector toolkit for marker-less integration of genes into Saccharomyces cerevisiae via CRISPR-Cas9.
Jessop-Fabre MM, Jakociunas T, Stovicek V, Dai Z, Jensen MK, Keasling JD, Borodina I
Biotechnol J. 2016 Aug;11(8):1110-7. doi: 10.1002/biot.201600147. Epub 2016 Jun 23.
PubMed Article

Plasmids from Article

ID Plasmid Purpose  
73271pCfB2899(X-2 MarkerFree)EasyClone-MarkerFree backbone vector for the addition of 1 or 2 genes and promoters for integration into site X-2 (Chr X: 194944..195980)
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73272pCfB3034(X-3 MarkerFree)EasyClone-MarkerFree backbone vector for the addition of 1 or 2 genes and promoters for integration into site X-3 (Chr X: 223616..224744)
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73273pCfB3035(X-4 MarkerFree)EasyClone-MarkerFree backbone vector for the addition of 1 or 2 genes and promoters for integration into site X-4 (Chr X: 236336..237310)
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73274pCfB3036(XI-1 MarkerFree)EasyClone-MarkerFree backbone vector for the addition of 1 or 2 genes and promoters for integration into site XI-1 (Chr XI: 67491..68573)
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73275pCfB2903(XI-2 MarkerFree)EasyClone-MarkerFree backbone vector for the addition of 1 or 2 genes and promoters for integration into site XI-2 (Chr XI: 91575..92913)
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73276pCfB2904(XI-3 MarkerFree)EasyClone-MarkerFree backbone vector for the addition of 1 or 2 genes and promoters for integration into site XI-3 (Chr XI: 93378..94567)
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73277pCfB3037(XI-5 MarkerFree)EasyClone-MarkerFree backbone vector for the addition of 1 or 2 genes and promoters for integration into site XI-5 (Chr XI: 11779..118967)
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73278pCfB3038(XII-1 MarkerFree)EasyClone-MarkerFree backbone vector for the addition of 1 or 2 genes and promoters for integration into site XII-1 (Chr XII: 795787..796720)
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73279pCfB3039(XII-2 MarkerFree)EasyClone-MarkerFree backbone vector for the addition of 1 or 2 genes and promoters for integration into site XII-2 (Chr XII: 808805..809939)
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73280pCfB3040(XII-4 MarkerFree)EasyClone-MarkerFree backbone vector for the addition of 1 or 2 genes and promoters for integration into site XII-4 (Chr XII:830227..831248)
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73281pCfB2909(XII-5 MarkerFree)EasyClone-MarkerFree backbone vector for the addition of 1 or 2 genes and promoters for integration into site XII-5 (Chr XII: 839226..840357)
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73282pCfB3020(gRNA X-2)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at site X-2
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73283pCfB3041(gRNA X-3)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at site X-3
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73284pCfB3042(gRNA X-4)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at site X-2
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73285pCfB3043(gRNA XI-1)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at site XI-1
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73286pCfB3044(gRNA XI-2)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at site XI-2
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73287pCfB3045(gRNA XI-3)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at site XI-3
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73288pCfB3046(gRNA XI-5)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at site XI-5
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73289pCfB3047(gRNA XII-1)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at site XII-1
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73290pCfB3048(gRNA XII-2)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at site XII-2
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73291pCfB3049(gRNA XII-4)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at site XII-4
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73292pCfB3050(gRNA XII-5)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at site XII-5
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73293pCfB3051(gRNA X-3 XI-2 XII-2)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at sites X-3, XI-2, and XII-2
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73294pCfB3052(gRNA X-4, XI-3, XII-5)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at sites X-4, XI-3, and XII-5
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73295pCfB3053(gRNA X-2, XI-5, XII-4)EasyClone-MarkerFree guiding RNA vector to direct Cas9 to cut at sites X-2, XI-5, and XII-4
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73296pCfB1975 (pUC19-PTEF1-PPGK1)Example plasmid containing a bidirectional promoter
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78230p1977 (pUC19-PTDH3-PTEF1)Example plasmid containing a bidirectional promoter of pTDH3 and pTEF1
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78231pCfB2312 (TEF1p-Cas9-CYC1t_kanMX)Expresses Cas9 protein, under the control of the TEF1 promoter, and CYC terminator. KanMX resistance
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78232pTAJAK-71 (pESC-NatMXsyn-USER)Empty vector, for the insertion of gRNA expression cassettes into the vector.
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