LOCUS Exported 3922 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 pGDP1 NDM-1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3922) 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 3922) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3922 /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 1180..1197 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 1259..1286 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" CDS complement(1377..2192) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 2100..2119 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2184..2203 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 2314..2902 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2803..2822 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3056..3073 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3088..3227 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3153..3172) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3313..3335 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3329..3520) /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 atggaactgc cgaacattat gcacccggtt gcgaaactga 361 gcacggccct ggcagcagcc ctgatgctga gcggttgtat gccgggtgaa attcgtccga 421 ccatcggcca gcaaatggaa acgggcgatc agcgttttgg tgacctggtg ttccgtcagc 481 tggcaccgaa cgtttggcaa catacctcat atctggatat gccgggcttt ggtgcagttg 541 cttcgaatgg tctgatcgtt cgtgatggtg gtcgtgtcct ggtggttgac accgcatgga 601 cggatgacca gaccgcccaa attctgaact ggatcaaaca ggaaattaat ctgccggtcg 661 cactggcagt cgtgacccat gcacaccaag ataaaatggg tggtatggac gcactgcatg 721 cagcaggcat tgctacgtat gcaaacgctc tgtccaatca gctggcgccg caagaaggta 781 tggtggcagc tcagcacagc ctgacctttg cagcaaacgg ttgggtcgaa ccggcaacgg 841 ccccgaattt tggtccgctg aaagtgttct acccgggccc gggtcacacc agtgataaca 901 tcacggttgg cattgatggt accgacatcg ccttcggcgg ttgcctgatt aaagatagta 961 aagcgaaatc cctgggcaat ctgggtgatg ccgacacgga acattacgca gctagcgcac 1021 gtgcttttgg tgcagcattc ccgaaagcat ctatgatcgt gatgagccac tctgcaccgg 1081 attcacgcgc tgcgattacc cacaccgccc gtatggctga caaactgcgt tgaggatccg 1141 aattcgagct ccgtcgacaa gcttgcggcc gcactcgagc accaccacca ccaccactga 1201 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 1261 aaggccatcc gtcaggatgg ccttctggtg gcacttttcg gggaaatgtg cgcggaaccc 1321 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag 1381 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca 1441 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg 1501 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt 1561 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa 1621 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca 1681 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc 1741 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc 1801 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct 1861 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca 1921 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt 1981 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac 2041 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta 2101 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta 2161 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa 2221 gcagacagtt ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 2281 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 2341 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 2401 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 2461 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 2521 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 2581 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 2641 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 2701 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 2761 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 2821 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 2881 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 2941 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 3001 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga 3061 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg 3121 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 3181 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc 3241 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag 3301 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg 3361 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct 3421 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa 3481 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg 3541 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg 3601 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat 3661 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag 3721 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg 3781 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc 3841 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta 3901 tcggtgattc attctgctaa cc //