LOCUS Exported 1018 bp ds-DNA linear SYN 18-MAY-2021 DEFINITION EasyClone-MarkerFree backbone vector for the addition of 1 or 2 genes and promoters for integration into site X-3 (Chr X: 223616..224744). ACCESSION . VERSION . KEYWORDS pCfB3034(X-3 MarkerFree) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 1018) AUTHORS Jessop-Fabre MM, Jakociunas T, Stovicek V, Dai Z, Jensen MK, Keasling JD, Borodina I TITLE EasyClone-MarkerFree: A vector toolkit for marker-less integration of genes into Saccharomyces cerevisiae via CRISPR-Cas9. JOURNAL Biotechnol J. 2016 Aug;11(8):1110-7. doi: 10.1002/biot.201600147. Epub 2016 Jun 23. PUBMED 27166612 REFERENCE 2 (bases 1 to 1018) AUTHORS . TITLE Direct Submission JOURNAL Exported May 18, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..1018 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 217..250 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" primer_bind 879..901 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 893..910 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 894..910 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 920..939 /label=T7 /note="T7 promoter, forward primer" promoter 920..938 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 tctattattt atgatttttt gtatatacaa tatttctagt ttgttaaaga gaattaagaa 61 aataaatctc gaaaataata aagggaaaat cagtttttga tatcaaaatt atacatgtca 121 acgataatac aaaatataat acaaactata agatgttatc agtatttatt atgcatttag 181 aataaatttt gtgtcgccct tattcgactc actatagaag ttcctattct ctagaaagta 241 taggaacttc acttcatttt atttaaattt gcggccgcgc tgaggtctta atgaggtggt 301 tattgatcac cggaaagttt ggcattaaaa acaaacgaca gcacaaagga actttcaccc 361 tacagcacct tctgtgtgca tgattacatg tacccgagta gcgaaaaaag ccagaaaatg 421 tgcaagaata aaggtatctc aatgaacgag ctcgagcgtc aagagacggg tataagcgaa 481 gaagcgcaat gccgccagcc aagcaaagcc aaacgaagag atcgaccttc catggggtcg 541 caacttttcc cggtgacctc tacatgtgtg attgttagtt cagcgtaaca gagtttcaac 601 aacatgcata tacgagtgtg ccttcgtgcc tgattttggt tgattatccg gcttccaacc 661 tcgttaaatt ttcataaagt gcatatcgtg taatgtaaaa agtaaaaaga taaaaaagac 721 tcatatgaaa tcttgctgca tggagacaag tgcactcgaa ttctttactt atattttcgc 781 tcttgagctc gtccttttac tagcatatca atatccgttt cattgaaaag tggattaatg 841 cctcagcact agtcctgcag gggtaacgcc agggttttcc cagtcacgac gttgtaaaac 901 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggtc gcgatcttcg 961 agcgtcccaa aaccttctca agcaaggttt tcagtataat gttacatgcg tacacgcg //