LOCUS Exported 4355 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Part of GoldenGate Yarrowia lipolytica toolkit for assembling an expression plasmid for Y. lipolytic yeast. Promoter for gene expression in position 3 (P3_PGM). ACCESSION . VERSION . KEYWORDS GGE007 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4355) AUTHORS Larroude M, Park YK, Soudier P, Kubiak M, Nicaud JM, Rossignol T TITLE A modular Golden Gate toolkit for Yarrowia lipolytica synthetic biology. JOURNAL Microb Biotechnol. 2019 May 31. doi: 10.1111/1751-7915.13427. PUBMED 31148366 REFERENCE 2 (bases 1 to 4355) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4355 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 107..128 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 143..173 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 181..197 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 186..208 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 205..221 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 205..221 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 239..257 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 239..256 /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1243..1262) /label=T7 /note="T7 promoter, forward primer" promoter complement(1244..1262) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1269..1286) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1269..1285) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1278..1300) /label=M13/pUC Forward /note="In lacZ gene" CDS 1423..1725 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 1604..1623 /label=ccdB-fwd /note="ccdB gene, forward primer" CDS 2074..2868 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2128..2147) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2738..2757 /label=Neo-F /note="Neomycin resistance gene, forward primer" regulatory 3068..3077 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3074..3448 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" rep_origin 3586..4174 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4075..4094 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4328..4345 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat 241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca 301 ctagtaacgg ccgccagtgt gctggaattc gcccttggtc tctgtcatac caaccacaga 361 ttacgaccca ttcgcagtca cagttcacta gggtttgggt tgcatccgtt gagagtggtt 421 tgtttttaac cttctccatg tgctcactca ggttttgggt tcagatcaaa tcaaggcgtg 481 aaccactgtt tgaggacaaa tgtgacacaa ccaaccagtg tcaggggcaa gtccgtgaca 541 aaggggaaga tacaatgcaa ttactgacag ttacggactg cctcgatgcc ctaaccttgc 601 cccaaaataa gacaactgtc ctcgtttaag cgcaacccta ttcagcgtca cgtcataata 661 gcgtttggat agcactagtc tatgaggagc gttttatgtt gcggtgaggg cgattggtgc 721 tcatatgggt tcaattgagg tggtggaacg agcttagtct tcaattgagg tgcgagcgac 781 acaattgggt gtcacgtggc ctaattgacc tcggatcgtg gagtccccag ttatacagca 841 accacgaggt gcatgagtag gagacgtcac cagacaatag ggtttttttg gactggagag 901 ggtagggcaa aagcgctcaa cgggctgttt ggggagctat gggggaggaa ttggcgatat 961 ttgtgaggtt gacggctccg atttgcgtgt tttgtcgctt ctgcatctcc ccatacccat 1021 atcttccctc cccacctctt tccacgataa ttttacggat cagcaataag gttccttctc 1081 ctagtttcca cgtccatata tatctatgct gcgtcgtcct tttcgtgaca tcaccaaaac 1141 acatacaaaa ccacagagac caagggcgaa ttcaggcctg aattctgcag atatccatca 1201 cactggcggc cgctcgagca tgcatctaga gggcccaatt cgccctatag tgagtcgtat 1261 tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa 1321 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc 1381 accgatcgcc cttcccaaca gttgcgcagc ctatacgtac ggcagtttaa ggtttacacc 1441 tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg 1501 ccggggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc 1561 cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat 1621 atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa 1681 aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc aggcatgaga 1741 ttatcaaaaa ggatcttcac ctagatcctt ttcacgtaga aagccagtcc gcagaaacgg 1801 tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca 1861 aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta 1921 tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc 1981 tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaagc 2041 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 2101 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 2161 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 2221 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg 2281 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 2341 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 2401 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 2461 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 2521 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 2581 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc 2641 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 2701 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 2761 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 2821 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat tattaacgct 2881 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcatc 2941 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca 3001 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatagcacgt 3061 gaggagggcc accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt 3121 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga 3181 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca 3241 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc 3301 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga 3361 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt 3421 gcacttcgtg gccgaggagc aggactgaca cgtgctaaaa cttcattttt aatttaaaag 3481 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc 3541 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt 3601 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt 3661 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat 3721 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 3781 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 3841 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 3901 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 3961 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 4021 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa 4081 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt 4141 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 4201 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc 4261 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac 4321 cgagcgcagc gagtcagtga gcgaggaagc ggaag //