LOCUS Exported 2726 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Level 1 cloning vector. ACCESSION . VERSION . KEYWORDS pUG-P5 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 143..164 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 179..209 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 217..233 /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 222..244 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 241..257 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 241..257 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 253..576 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS DAA" misc_feature 270..326 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(327..344) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(327..343) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(336..358) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(552..571) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(711..729) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 797..901 /gene="bla" /label=AmpR promoter CDS 902..1762 /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(1120..1139) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1933..2521 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2422..2441 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2675..2692 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 tccccgcgcg ttggccgatt cattaatcac tctgtggttc agaagacatc agattaactt 61 taagaaggag atatacaatg ggagaccgca gctggcacga caggtttccc gactggaaag 121 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt 181 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca 241 caggaaacag ctatgaccat gattacgcca agcttgcatg cctgcaggtc gactctagag 301 gatccccggg taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa 361 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt 421 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa 481 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg 541 tgcactctca gtacaatctg ctctgatgcc gcataaggtc tcagctttgt gttgtcttct 601 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc 661 ggcatccgct tacagacaag ctgtgaccca cgaagtgagt ggacgaaagg gcctcgtgat 721 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac 781 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 841 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 901 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc 961 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc 1021 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc 1081 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc 1141 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt 1201 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt 1261 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat 1321 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct 1381 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat 1441 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc 1501 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg 1561 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggttc 1621 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta 1681 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc 1741 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga 1801 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 1861 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 1921 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 1981 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 2041 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 2101 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 2161 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 2221 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 2281 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 2341 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 2401 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 2461 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 2521 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 2581 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc 2641 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga 2701 tgagcgccca atacgcaaac cgcctc //