LOCUS Exported 3320 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Cloning Module. ACCESSION . VERSION . KEYWORDS pUG41780 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3320) AUTHORS Kowarschik K, Hoehenwarter W, Marillonnet S, Trujillo M TITLE UbiGate: a synthetic biology toolbox to analyse ubiquitination. JOURNAL New Phytol. 2018 Mar;217(4):1749-1763. doi: 10.1111/nph.14900. Epub 2017 Nov 30. PUBMED 29194629 REFERENCE 2 (bases 1 to 3320) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3320 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(334..438) /gene="bla" /label=AmpR promoter primer_bind 506..524 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(562..584) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 684..703 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 897..919 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 911..928 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 912..928 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1480..1496) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1480..1496) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1493..1515) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1504..1520 /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(1528..1558) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1573..1594 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1711..1728) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1882..2470) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1962..1981) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2641..112) /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 ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc 61 caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc atgatgttta 121 actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctccat aacatcaaac 181 atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtaccccaa 241 aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc 301 ggtcaaggtt ctggaccagt tgcgtgagcg catactcttc ctttttcaat attattgaag 361 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 421 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat 481 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gactcgcgcg 541 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagatgg tcacagcttg 601 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg 661 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat 721 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc 781 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca 841 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca 901 gtcacgacgt tgtaaaacga cggccagtga attcgagctc ggtacccttt gaagacaaca 961 gagaggatgc acatgtgacc gagggattgt cttcgtggaa acttgggact tcagattcca 1021 ccccttgcaa gcgtggaaag ccttccgacc aagggaagtg tcggatgtag aggaaatgga 1081 aagtttgttc tcagatgggg acttgcttga ttgcttcaca agaatgccag cttatgcggt 1141 aaacgcagag gaagatttag ctgcaatcag gaaaacgccc gagatggatg tcggtcaaga 1201 agttaaagaa cctgcaggag acagaaatca atactcaaac cctgcagaaa ctttcctcaa 1261 caagctccac aggaaacaca gtagggaggt gaaacaccag gccgcaaaga aagctaaacg 1321 cctagctgaa atccaggagt caatgagagc tgaaggtgat gccgaaccaa atgaaataag 1381 cgggggcatg ggggcaatac ccagcaacgc cgaacttcct gggcccaaag gggatcctct 1441 agagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 1501 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 1561 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 1621 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 1681 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 1741 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 1801 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 1861 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 1921 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 1981 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 2041 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 2101 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 2161 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 2221 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 2281 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 2341 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 2401 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 2461 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 2521 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 2581 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 2641 ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg 2701 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag 2761 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt 2821 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt 2881 tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc 2941 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa 3001 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc 3061 tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg 3121 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc 3181 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc 3241 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg 3301 tggcttcagg ccgccatcca //