LOCUS Exported 4036 bp ds-DNA circular SYN 13-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 aph(3'')-Ia SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4036) 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 4036) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4036 /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 1161..1178 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 1240..1267 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 1292..1396 /gene="bla" /label=AmpR promoter CDS 1397..2257 /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(1615..1634) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2428..3016 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2917..2936 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3170..3187 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3202..3341 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3267..3286) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3427..3449 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3443..3634) /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 atgagtgatc accccgggcc gggggccgtc acgccggagc 361 tgttcggcgt gggcggcgac tggctggccg tcaccgcggg cgaatcgggc gcctccgtct 421 ttcgcgccgc ggacgccacc cggtacgcca agtgcgtgcc cgccgcggac gcggccggtc 481 ttgaggcgga acgcgaccgg atcgcctggc tgagcgggca gggcgtaccg ggcccccgcg 541 tcctcgactg gtacaccggt gacgcgggcg cctgcctggt cacccgtgcc gtccccggcg 601 tacccgctga tcgggtgggc gccgatgacc ttcgcactgc ctggggggcc gtcgcggacg 661 cggtccgtcg gctgcacgag gtacccgtgg cctcgtgtcc gttccgccgg gggctggact 721 ccgtggtcga cgccgcccgt gacgtggcgg cccgtggcgc ggtgcatccg gagttcctgc 781 cggtggagca gcggctcgct cccccggcgg agctgctggc ccggctcacc ggggagctcg 841 cccgtcggcg cgatcaggag gccgccgaca cggtcgtctg ccacggtgat ctctgcctgc 901 ccaacatcgt cctccatccg gagaccctgg aggtgtcggg cttcatcgac ctgggacggc 961 tcggggcggc cgaccgccac gccgacctgg cgctgctgct ggccaacgcg cgcgagacct 1021 gggtggacga ggagcgggcg cggttcgccg acgcggcgtt cgccgagcgc tacggcatcg 1081 ccccggaccc ggaacggctg cgcttctacc tccatctcga tccgctcacc tggggctaga 1141 agcttgcggc cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag 1201 cccgaaagga agctgagttg gctgctgcca ccgctgagca aaaggccatc cgtcaggatg 1261 gccttctggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 1321 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 1381 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 1441 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 1501 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 1561 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 1621 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 1681 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 1741 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 1801 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 1861 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 1921 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga 1981 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 2041 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 2101 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 2161 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 2221 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 2281 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 2341 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 2401 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 2461 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 2521 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct 2581 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 2641 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 2701 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 2761 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 2821 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 2881 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 2941 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 3001 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 3061 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 3121 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 3181 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat 3241 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag 3301 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac 3361 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga 3421 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc 3481 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat 3541 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg 3601 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc 3661 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg 3721 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc 3781 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag 3841 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg 3901 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg 3961 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg 4021 attcattctg ctaacc //