LOCUS Exported 3318 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Cloning Module. ACCESSION . VERSION . KEYWORDS pUG41822 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3318) 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 3318) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3318 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(51..67) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(51..67) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(64..86) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 75..91 /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(99..129) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 144..165 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(282..299) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(453..1041) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(533..552) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1212..2003) /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" promoter complement(2223..2327) /gene="bla" /label=AmpR promoter primer_bind 2395..2413 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(2451..2473) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2573..2592 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2786..2808 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2800..2817 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2801..2817 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg 61 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 121 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 181 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 241 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 301 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 361 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 421 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 481 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 541 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 601 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 661 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 721 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 781 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 841 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta 901 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 961 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 1021 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 1081 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 1141 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 1201 tggtctgaca gttatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat 1261 tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt 1321 ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca 1381 gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta 1441 agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt 1501 tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct 1561 ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg 1621 tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat 1681 tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg 1741 acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg 1801 ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca 1861 atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc 1921 aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact 1981 tcggcgatca ccgcttccct catgatgttt aactttgttt tagggcgact gccctgctgc 2041 gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc 2101 ttggatgccc gaggcataga ctgtacccca aaaaaacagt cataacaagc catgaaaacc 2161 gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc 2221 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 2281 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 2341 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 2401 cgtatcacga ggccctttcg actcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 2461 atgcagctcc cggagatggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc 2521 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca 2581 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg 2641 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga 2701 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 2761 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa 2821 ttcgagctcg gtaccctttg aagacaagag cgaggatgca catgtgaccg agggattgtc 2881 ttcgtggaaa cttgggactt cagattccac cccttgcaag cgtggaaagc cttccgacca 2941 agggaagtgt cggatgtaga ggaaatggaa agtttgttct cagatgggga cttgcttgat 3001 tgcttcacaa gaatgccagc ttatgcggta aacgcagagg aagatttagc tgcaatcagg 3061 aaaacgcccg agatggatgt cggtcaagaa gttaaagaac ctgcaggaga cagaaatcaa 3121 tactcaaacc ctgcagaaac tttcctcaac aagctccaca ggaaacacag tagggaggtg 3181 aaacaccagg ccgcaaagaa agctaaacgc ctagctgaaa tccaggagtc aatgagagct 3241 gaaggtgatg ccgaaccaaa tgaaataagc gggggcatgg gggcaatacc cagcaacgcc 3301 gaacttcctg ggcccaaa //