LOCUS Exported 4421 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION GoldenBraid vector pDGB2-Omega-1, adapted for S. cerevisiae genome integration by flanking the GB cassette with recombination arms of homology for integration at the YPRC-Delta-15 solo LTR loci. ACCESSION . VERSION . KEYWORDS YPRC-Delta-15-Omega-1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4421) AUTHORS Perez-Gonzalez A, Kniewel R, Veldhuizen M, Verma HK, Navarro-Rodriguez M, Rubio LM, Caro E TITLE Adaptation of the GoldenBraid modular cloning system and creation of a toolkit for the expression of heterologous proteins in yeast mitochondria. JOURNAL BMC Biotechnol. 2017 Nov 13;17(1):80. doi: 10.1186/s12896-017-0393-y. PUBMED 29132331 REFERENCE 2 (bases 1 to 4421) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4421 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(1147..1470) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS DAA" primer_bind 1152..1171 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1365..1387 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1379..1396 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1380..1396 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" misc_feature 1397..1453 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(1466..1482) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1466..1482) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1479..1501) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1490..1506 /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)." promoter complement(1514..1544) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1559..1580 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(2697..3285) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2777..2796) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3373..4164) /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" ORIGIN 1 atgtccactg ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg 61 gtagcagcga agtcgaggca tttctgtcct ggtgggcggc cgctagggat aacagggtaa 121 tcgagatagt ttgacgttcg ttttttactt tgaatatact cgtagtcttt ttactttttg 181 agtataaaaa aaaaatgact agcaaaataa aattagtagt ctaaaaaaga aagctcgcac 241 tcaggatcga actaaggacc aacagatttg caatctgctg cgctaccact gcgccatacg 301 agcttttgaa ttatggtaat tttgattatc ctagaatgtt atatctcaat atctcaatat 361 attttggaca tctatgaaac acccataaag cagccgctac caaacagaca agattcagta 421 tgtaaggtaa ataccttttt gcacagttaa actacccaaa cttattaaag cttgataaat 481 tactgaaatt ccacctttca gttagattca ggcctcatat agattagata tagggtacgt 541 aacattctgt caaccaagtt gttggaatga aagtctaaaa tgtcatctat tcggtagcac 601 tcatgttact agtatactgt cacatgcggt gtaacgtggg gacataaaac agacatcaaa 661 tataatggaa gctgaaatgc aaagatcgat aatgtaatag gaatgaaaca tataaaacga 721 aaggagaagt aatggtaata ttagtatgta gaaataccga ttcaattttg gggattctta 781 tattctcgag agaatttcta gtataatctg tatacataat attataggct ttaccaacaa 841 tggaatttcg acaattatca tattattcac caattaatca caagttggta atgagtttga 901 taacaagtta ctttcttaac aacgttagta tcgtcaaaac actcggtttt actcgagctt 961 gtagcacaat aataccgtgt agagttctgt attgttcttc ttagtgcttg tatatgctca 1021 tcccgacctt ccattggatc cggtctcagg agtgagacgc acagcttgtc tgtaagcgga 1081 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 1141 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 1201 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 1261 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 1321 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 1381 taaaacgacg gccagtgaat tcgagctcgg tacccgggga tcctctagag tcgacctgca 1441 ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 1501 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 1561 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 1621 tgtcgtgcca gctgcgtctc acgctgtcat gagaccggat cctttgcgaa accctatgct 1681 ctgttgttcg gatttgaaat tttaaaacta cattaatgtg ttagtttttc tttctttctt 1741 tctttgtctt gacgtgattt ggacttctgt cttgcattcg cgtccattca tctgacccaa 1801 tattcctttt ggttttgtta tccttataaa aagaaaggaa gcttcttaga gggaaaaaaa 1861 tgatgaagag taatgccaaa atataaataa ataaataaat atgaaaatca ttttctattt 1921 ttaatagaat aagaagagca tcttaagatt acaatttcaa gaaatagttt acacagtata 1981 tccaataact ccaataaact actttcctat acaaatttct atggtgggat taatagtaaa 2041 acttctgtac ttctctaatt caccaagaaa ttaaggtaaa catctggtaa gcactatcca 2101 gctttttgct attacacata tggcttttct gcaatcattt cttcccattt tgtctcaagc 2161 cgttagtctt gaaaccacag gcggagtaga gttacttgat gcggtatttt acatgccttt 2221 tttcactgca aaaaaaatga aatacatatt tacacgattt gcaggacagt ttacgatagt 2281 gagtatgcag aatagttaac acctttgttt tatccttttg tgtcttaatt atatgatata 2341 aaggcgcctg gcgttatcgg atagtaatag atgctagtta tcaacatttc acaattgaag 2401 gaaataaagt tgaagtactc aacaaaaact tacttcagaa ttaaattttt ggggggaaca 2461 taggcatcct atgacaggtg accacaagcc cctcaacgca atctaatatt ttacaaagtg 2521 gtaaaattct ttcgttcttc gttttaatat acagtcattt attgattcta ttacattaat 2581 attcctacgc ttcggctcac ataattaaca ggacttcgag tccgttaaac ttgggatcaa 2641 ctaatttcta cggattaggg ataacagggt aatgcggccg ccgcgttgct ggcgtttttc 2701 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 2761 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 2821 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 2881 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 2941 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 3001 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 3061 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 3121 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc 3181 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 3241 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 3301 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 3361 cattctaggt gattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa 3421 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg 3481 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc 3541 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt 3601 aagcactaca tttcgctcat caccagccca gtcgggcggc gagttccata gcgttaaggt 3661 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc 3721 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat 3781 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa 3841 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt 3901 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc 3961 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc 4021 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag 4081 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac 4141 ttcggcgatc accgcttccc tcataatgtt taactttgtt ttagggcgac tgccctgctg 4201 cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg 4261 cttggatgcc cgaggcatag actgtacccc aaaaaaacag tcataacaag ccatgaaaac 4321 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg 4381 catacgctac ttgcattaca gcttacgaac cgaacaggct t //