LOCUS Exported 2927 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION empty BB3 for insertion and overexpression of 5 genes in P. pastoris. ACCESSION . VERSION . KEYWORDS BB3aK_AF SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2927) AUTHORS Prielhofer R, Barrero JJ, Steuer S, Gassler T, Zahrl R, Baumann K, Sauer M, Mattanovich D, Gasser B, Marx H TITLE GoldenPiCS: a Golden Gate-derived modular cloning system for applied synthetic biology in the yeast Pichia pastoris. JOURNAL BMC Syst Biol. 2017 Dec 8;11(1):123. doi: 10.1186/s12918-017-0492-3. PUBMED 29221460 REFERENCE 2 (bases 1 to 2927) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2927 /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 761..2117 /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" promoter 761..1104 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 1105..1914 /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(1172..1191) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" terminator 1920..2117 /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 ttctcatagc 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 agaacagtat 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 gcgatcggag accggatccc tcgagggtct ccgcttcatg gacatggagg cccagaatac 781 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat 841 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc 901 gcgaagcaaa aattacggct cctcgctgca gacgtgcgag cagggaaacg ctcccctcac 961 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt 1021 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca 1081 catcacatcc gaacataaac aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga 1141 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg 1201 caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg 1261 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg 1321 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca 1381 tggttactca ccactgcgat ccccggcaaa acagcattcc aggtattaga agaatatcct 1441 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt 1501 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca 1561 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct 1621 gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc 1681 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt 1741 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac 1801 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat 1861 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagta 1921 ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg 1981 tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca 2041 tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa 2101 tgctggtcgc tatactggct tcagcaacgt tgtcactgaa gttggcatca gtatctacaa 2161 accctacata cctcttgaag gtccagaagg taagcatttg cccgctccat tcctacccga 2221 catatccact ctacatttta tagttcaagg caccgaaaaa gttcgaaaca agaagtttgt 2281 tcctgataac aaggatttct ttattggtgg gacttcattt actgtctcca agaaggacat 2341 ttctgctgtc ataaccgaga tagtctcaca gttcgagtct actgatgacc aaaagtcaga 2401 gaaatttacc atgaaatctc ctcctccggt tgctttagtt gggcataatc tgataggagg 2461 cctcaagaca ctgaagaatg cgggtatcac cattcccata cttccaatca taggatccgg 2521 cgcgccgata ctcgagaatt atggcttaat caagtgaata catcaaagtc aaacttaaaa 2581 tacattcttc gcaggcttgg cttgccacat agttttcttc acaatgctgc aaatgacgct 2641 tattataccc ttttggcatc gttgaagctt gcacaaacga acgtcccact taatcttctg 2701 tactctgaag aggagtggga aataccaaga aaaacatcaa actcgaatga ttttcccgaa 2761 cccctaccac aagatattca tcagctgcga gataggctga tcaggagcaa gctcgtacga 2821 gaagaaacaa aatgacaaaa aaaatcctat actatatagg ttacaaataa aaaagtatca 2881 aaaatgaagc ctgcatctct caggcaaatg gcattctgac atcctct //