LOCUS Exported 2642 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Plasmid for assembly of donor DNA templates consisting of three expression units. ACCESSION . VERSION . KEYWORDS BB3nK_ext_AD SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2642) AUTHORS Gassler T, Heistinger L, Mattanovich D, Gasser B, Prielhofer R. TITLE CRISPR/Cas9-mediated homology directed genome editing in Pichia pastoris JOURNAL (2018) In Press REFERENCE 2 (bases 1 to 2642) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2642 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(62..650) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(142..161) /label=pBR322ori-F /note="pBR322 origin, forward primer" gene 785..2141 /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" promoter 785..1128 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 1129..1938 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1196..1215) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" terminator 1944..2141 /label=TEF terminator /note="Ashbya gossypii TEF terminator" ORIGIN 1 cgctgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 61 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 241 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 481 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 661 tgatcttttc tacggggtgg agcgcaaaaa accccgcccc tgacagggcg gggttttttc 721 gcgatcgaag acccagctgg agaccctcga gggatccggt ctccgatcgg gtcttcagct 781 catggacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc 841 atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc 901 catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacgtg 961 cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta 1021 gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa 1081 aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa 1141 aagactcacg tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat 1201 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag 1261 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca 1321 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat 1381 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca 1441 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg 1501 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta 1561 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt 1621 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg 1681 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt 1741 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac 1801 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg 1861 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg 1921 ctcgatgagt ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt 1981 catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt 2041 atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa 2101 tatcatgcgt caatcgtatg tgaatgctgg tcgctatact ggcttgtaag caatctggac 2161 aattatgtaa gcgggttacg taaacagtta tgtaagcaga aaaatttcaa acgacaaaac 2221 ttggggtcta cagacacagt agccagaaga ttgcactacc attcgactcc tcatgaccca 2281 ctctttcgat ccatgtagtt aggttaccgt ttttcctaat atttaaggat gttgaaaatt 2341 cattttcatt tttttcgttt ttaagatttt ctcacaactc ttccaaagat tactagttga 2401 cttttcaaat atttagggta tttttctcac tttttcctag caaactccaa ttggtgggtt 2461 cagtgcaatg gagtatcacc ttgcaaccac aacgtaatag ctaacttgtg gccaccatgt 2521 ctggttgtag agataattgg attctaatgt ggatcacatg actactcacg tgtcaaaaac 2581 ccaacctgac ttggcccagc ttagcaagaa tatttcgaat ccactcttgt ggcctagtgg 2641 ac //