LOCUS Exported 3320 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Cloning Module. ACCESSION . VERSION . KEYWORDS pUG41766 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" primer_bind 104..121 /label=L4440 /note="L4440 vector, forward primer" protein_bind 238..259 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 274..304 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 312..328 /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 317..339 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 336..352 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 336..352 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(904..921) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(904..920) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(913..935) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(1129..1148) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1248..1270 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1308..1326) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1394..1498 /gene="bla" /label=AmpR promoter CDS 1720..2511 /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" rep_origin 2682..3270 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3171..3190 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 61 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 121 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 181 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa 241 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat 301 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta 361 cgccaagctt gcatgcctgc aggtcgactc tagaggatcc cctttgaaga caattacgag 421 gatgcacatg tgaccgaggg attgtcttcg tggaaacttg ggacttcaga ttccacccct 481 tgcaagcgtg gaaagccttc cgaccaaggg aagtgtcgga tgtagaggaa atggaaagtt 541 tgttctcaga tggggacttg cttgattgct tcacaagaat gccagcttat gcggtaaacg 601 cagaggaaga tttagctgca atcaggaaaa cgcccgagat ggatgtcggt caagaagtta 661 aagaacctgc aggagacaga aatcaatact caaaccctgc agaaactttc ctcaacaagc 721 tccacaggaa acacagtagg gaggtgaaac accaggccgc aaagaaagct aaacgcctag 781 ctgaaatcca ggagtcaatg agagctgaag gtgatgccga accaaatgaa ataagcgggg 841 gcatgggggc aatacccagc aacgccgaac ttcctgggcc caaagggtac cgagctcgaa 901 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa 961 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga 1021 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct 1081 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc 1141 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg 1201 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accatctccg ggagctgcat 1261 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagt cgaaagggcc tcgtgatacg 1321 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt 1381 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 1441 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat 1501 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc 1561 ggttttcatg gcttgttatg actgtttttt tttggggtac agtctatgcc tcgggcatcc 1621 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt 1681 acgcagcagg gcagtcgccc taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc 1741 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg 1801 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat 1861 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc 1921 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa 1981 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg 2041 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc 2101 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt 2161 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat 2221 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg 2281 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc 2341 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct 2401 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa 2461 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata actgtcagac 2521 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 2581 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 2641 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 2701 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 2761 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 2821 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 2881 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 2941 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 3001 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 3061 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 3121 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 3181 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 3241 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 3301 ctggcctttt gctggccttt //