LOCUS Exported 2725 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Level 1 cloning vector. ACCESSION . VERSION . KEYWORDS pUG-TP6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2725) 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 2725) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2725 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 142..163 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 178..208 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 216..232 /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 221..243 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 240..256 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 240..256 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 252..575 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS DAA" misc_feature 269..325 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(326..343) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(326..342) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(335..357) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(551..570) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(710..728) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 796..900 /gene="bla" /label=AmpR promoter CDS 901..1761 /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(1119..1138) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1932..2520 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2421..2440 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2674..2691 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 tccccgcgcg ttggccgatt cattaatcac tctgtggttc agaagacatt gtgttaactt 61 taagaaggag atataccatg gagaccgcag ctggcacgac aggtttcccg actggaaagc 121 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt 181 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac 241 aggaaacagc tatgaccatg attacgccaa gcttgcatgc ctgcaggtcg actctagagg 301 atccccgggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa 361 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta 421 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat 481 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt 541 gcactctcag tacaatctgc tctgatgccg cataaggtct cagcttgagc ttgtcttcta 601 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg 661 gcatccgctt acagacaagc tgtgacccac gaagtgagtg gacgaaaggg cctcgtgata 721 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact 781 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 841 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 901 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 961 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca 1021 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 1081 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 1141 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 1201 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 1261 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc 1321 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 1381 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 1441 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct 1501 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 1561 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggttct 1621 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 1681 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 1741 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat 1801 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 1861 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 1921 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 1981 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 2041 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta 2101 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 2161 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 2221 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 2281 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 2341 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 2401 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 2461 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 2521 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 2581 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 2641 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggat 2701 gagcgcccaa tacgcaaacc gcctc //