LOCUS Exported 2725 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Level 1 cloning vector. ACCESSION . VERSION . KEYWORDS pUG-TP4 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 152..173 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 188..218 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 226..242 /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 231..253 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 250..266 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 250..266 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 262..585 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS DAA" misc_feature 279..335 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(336..353) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(336..352) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(345..367) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(561..580) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(720..738) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 806..910 /gene="bla" /label=AmpR promoter CDS 911..1771 /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(1129..1148) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1942..2530 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2431..2450 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2684..2701 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 aaaccgcctc tccccgcgcg ttggccgatt cattaatcac tctgtggttc agaagacatt 61 tacttaactt taagaaggag atataccatg gagaccgcag ctggcacgac aggtttcccg 121 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac 181 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac 241 aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgcatgc ctgcaggtcg 301 actctagagg atccccgggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt 361 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc 421 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg 481 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac 541 cgcatatggt gcactctcag tacaatctgc tctgatgccg cataaggtct cagcttcaga 601 ttgtcttcta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg 661 tctgctcccg gcatccgctt acagacaagc tgtgacccac gaagtgagtg gacgaaaggg 721 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc 781 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca 841 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa 901 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt 961 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca 1021 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag 1081 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc 1141 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca 1201 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt 1261 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct 1321 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt 1381 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga 1441 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact 1501 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc 1561 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga 1621 gcgtggttct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt 1681 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga 1741 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact 1801 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga 1861 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 1921 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 1981 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 2041 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta 2101 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 2161 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 2221 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 2281 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 2341 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 2401 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 2461 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 2521 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 2581 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 2641 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 2701 ggaagcggat gagcgcccaa tacgc //