LOCUS Exported 2893 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION empty BB3 for insertion and overexpression of 2 genes in P. pastoris. ACCESSION . VERSION . KEYWORDS BB3rN_AC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2893) 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 2893) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2893 /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..1880 /label=natMX6 /note="yeast selectable marker conferring nourseothricin resistance" promoter 761..1104 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 1105..1677 /codon_start=1 /gene="Streptomyces noursei nat1" /product="nourseothricin acetyltransferase" /label=NrsR /note="confers resistance to nourseothricin" /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT LCGLDTALYDGTASDGEQALYMSMPCP" terminator 1683..1880 /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 ccaattcatg gacatggagg cccagaatac 781 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat 841 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc 901 gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac 961 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt 1021 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca 1081 catcacatcc gaacataaac aaccatgggt accactcttg acgacacggc ttaccggtac 1141 cgcaccagtg tcccggggga cgccgaggcc atcgaggcac tggatgggtc cttcaccacc 1201 gacaccgttt ttcgcgtcac cgccaccggg gacggcttca ccctgcggga ggtgccggtg 1261 gacccgcccc tgaccaaggt gttccccgac gacgaatcgg acgacgaatc ggacgacggg 1321 gaggacggcg acccggactc ccggacgttc gtcgcgtacg gggacgacgg cgacctggcg 1381 ggcttcgtgg tcgtctcgta ctccggctgg aaccgccggc tgaccgtcga ggacatcgag 1441 gtcgccccgg agcaccgggg gcacggggtc gggcgcgcgt tgatggggct cgcgacggag 1501 ttcgcccgcg agcggggcgc cgggcacctc tggctggagg tcaccaacgt caacgcaccg 1561 gcgatccacg cgtaccggcg gatggggttc accctctgcg gcctggacac cgccctgtac 1621 gacggcaccg cctcggacgg cgagcaggcg ctctacatga gcatgccctg cccctaatca 1681 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata 1741 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca 1801 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt 1861 gaatgctggt cgctatactg gcttcatacc cagcattgac ctttggtatg agcatctgaa 1921 aaacaaccag gtgttgcaaa gttaaacatc cttctttgtt catatagaac ccactattca 1981 tggtactccc caatcgaatt tcacattctg gttttgaaat tacacaccac gttagcttat 2041 aagatttcat ataacttatt gatatacggt ttccattgtt cgaatagttg aggttgtatg 2101 taattcgatt gaaggggcca tttttgtttc ctacttttcc tgggagctta tccgatgcgc 2161 ttcaaagctg gaattgtaaa tatagagaaa aagaaggatg ttgttttatt cttgaaagag 2221 tataatttta cttctagcaa ctctcccact tcgcttgact tcatttattt cttgggcaca 2281 taggcgtagt aatctagacc aacagataat ttgccggaat gatatagcga ttggaaaatg 2341 aactgaaatt ttttgctgtc tttcaatttg acgggcagtt catcaggcgc gccgtgaccg 2401 accatataaa tacgttgaga atgttattct tcctcgtagt tgaagtggct tcataatttc 2461 agaactcaat agataaacta ggatgtttta aagcaattaa tgctcacaag taaggagcga 2521 ctctcttgct tttcgaatac taaaagtatc gtcccaaccc agaaaaaaag acctcttaac 2581 tgcaaaataa actctatata tttcttctaa aacagtttca ggttggatag tatcgcattc 2641 tcatcacttc taactagtag gccatgagat atattaacgt ttacttgagt tctaagttct 2701 ccgaattaga tgcacagcac aaacaagatt aggtttcact tggtacaaaa tacgaacaga 2761 gtttaaggtc gtaatttcat ttcgttattg atccccacaa tctattctta tcacagtcat 2821 cagatagtcg cgaaaaagca tgcagaaaag ggggtcgtcc ctatctaagt tgtagcatta 2881 caacaaatat gac //