LOCUS Exported 3566 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION This plasmid is part of the Minimal Antibiotic Resistance Platform (ARP) Kit (Addgene #1000000143). Low copy-number plasmid that constitutively expresses genes using the Pbla promoter.. ACCESSION . VERSION . KEYWORDS pGDP3 rmtB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3566) AUTHORS Cox G, Sieron A, King AM, De Pascale G, Pawlowski AC, Koteva K, Wright GD TITLE A Common Platform for Antibiotic Dereplication and Adjuvant Discovery. JOURNAL Cell Chem Biol. 2017 Jan 19;24(1):98-109. doi: 10.1016/j.chembiol.2016.11.011. Epub 2016 Dec 22. PUBMED 28017602 REFERENCE 2 (bases 1 to 3566) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3566 /organism="synthetic DNA construct" /mol_type="other DNA" RBS 232..254 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 273..290 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 300..317 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 691..708 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 770..797 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 822..926 /gene="bla" /label=AmpR promoter CDS 927..1787 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1145..1164) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1958..2546 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2447..2466 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2700..2717 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2732..2871 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(2797..2816) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2957..2979 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(2973..3164) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" ORIGIN 1 agtaaggcaa ccccgccagc ctagccggga gatctggcct attggttaaa aaatgagctg 61 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca 121 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 181 tgtatccgct catgagacaa taaccctgat aaatgcttct ctagaaataa ttttgtttaa 241 ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac agcagcggcc 301 tggtgccgcg cggcagccat atggctagca tgaacatcaa cgatgccctc acctccatcc 361 tggcctcaaa aaaataccgc gccctttgcc ctgcaggatg tgctgtgtgc gccgcccgcc 421 gaagccggcg acctggcgct gatttttaag cttttgcccc tgctggagcg ggagcaggcc 481 ggttctgcca tggcactttt acaatccctc aataccccgc gcatggctgt cagctttccc 541 acgcgtagtt taggcgggcg tggaaaaggc atggaggcga actacgccgc atggttcgag 601 ggcggcttgc ccgccgagtt tgagattgag gataaaaaga ccatcggaac agaacttata 661 tacttgataa aaaagaatgg ataactcgag caccaccacc accaccactg agatccggct 721 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca aaaggccatc 781 cgtcaggatg gccttctggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 841 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 901 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc 961 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 1021 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg 1081 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag 1141 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc 1201 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta 1261 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg 1321 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca 1381 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 1441 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat 1501 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg 1561 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata 1621 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta 1681 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 1741 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag 1801 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 1861 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 1921 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 1981 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 2041 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 2101 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 2161 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 2221 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 2281 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 2341 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 2401 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 2461 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 2521 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 2581 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 2641 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca 2701 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc 2761 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata 2821 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac 2881 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga 2941 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa 3001 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct 3061 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg 3121 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa 3181 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata 3241 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg 3301 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat 3361 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata 3421 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc 3481 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg 3541 cgtatcggtg attcattctg ctaacc //