LOCUS Exported 2726 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Level 1 cloning vector. ACCESSION . VERSION . KEYWORDS pUG-P3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2726) 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 2726) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2726 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 106..127 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 142..172 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 180..196 /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 185..207 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 204..220 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 204..220 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 216..539 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS DAA" misc_feature 233..289 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(290..307) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(290..306) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(299..321) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(515..534) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(674..692) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 760..864 /gene="bla" /label=AmpR promoter CDS 865..1725 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1083..1102) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1896..2484 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2385..2404 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2638..2655 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 ttcagaagac atactattaa ctttaagaag gagatataca atgggagacc gcagctggca 61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct 121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat 181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc 241 atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg 301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac 361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac 421 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt 481 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcataag 541 gtctcagctt ttacttgtct tctaagccag ccccgacacc cgccaacacc cgctgacgcg 601 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac ccacgaagtg 661 agtggacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg 721 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta 781 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt 841 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc 901 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa 961 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt 1021 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt 1081 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc 1141 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg 1201 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg 1261 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac 1321 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca 1381 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta 1441 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat 1501 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa 1561 tctggagccg gtgagcgtgg ttctcgcggt atcattgcag cactggggcc agatggtaag 1621 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat 1681 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt 1741 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg 1801 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga 1861 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 1921 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 1981 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 2041 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 2101 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 2161 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 2221 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 2281 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 2341 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 2401 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 2461 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 2521 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac 2581 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc 2641 gagtcagtga gcgaggaagc ggatgagcgc ccaatacgca aaccgcctct ccccgcgcgt 2701 tggccgattc attaatcact ctgtgg //