LOCUS Exported 4588 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 pLac promoter.. ACCESSION . VERSION . KEYWORDS pGDP2 MCR-1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4588) 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 4588) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4588 /organism="synthetic DNA construct" /mol_type="other DNA" RBS 367..389 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 2028..2045 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 2107..2134 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" CDS complement(2225..3040) /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 2948..2967 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3032..3051 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 3162..3750 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3651..3670 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3904..3921 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3936..4075 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4001..4020) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4161..4183 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(4177..4368) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" ORIGIN 1 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc 61 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca 121 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa 181 ccagtaaggc aaccccgcca gcctagccgg gagatctgcg gcggcggtgc tcaacggcct 241 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc 301 cggacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggtctaga 361 aataattttg tttaacttta agaaggagat ataccatggt ccagcatact cctgtgtggt 421 accgacgctc ggtcagtccg tttgttcttg tggcgagtgt tgccgttttc ttgaccgcga 481 ccgccaatct tacctttttt gataaaatca gccaaaccta tcccatcgcg gacaatctcg 541 gctttgtgct gacgatcgct gtcgtgctct ttggcgcgat gctactgatc accacgctgt 601 tatcatcgta tcgctatgtg ctaaagcctg tgttgatttt gctattaatc atgggcgcgg 661 tgaccagtta ttttactgac acttatggca cggtctatga tacgaccatg ctccaaaatg 721 ccctacagac cgaccaagcc gagaccaagg atctattaaa cgcagcgttt atcatgcgta 781 tcattggttt gggtgtgcta ccaagtttgc ttgtggcttt tgttaaggtg gattatccga 841 cttggggcaa gggtttgatg cgccgattgg gcttgatcgt ggcaagtctt gcgctgattt 901 tactgcctgt ggtggcgttc agcagtcata tggccagttt ctttcgcgtg cataagccgc 961 tgcgtagcta tgtcaatccg atcatgccaa tctactcggt gggtaagctt gccagtattg 1021 agtataaaaa agccagtgcg ccaaaagata ccatttatca cgccaaagac gcggtacaag 1081 caaccaagcc tgatatgcgt aagccacgcc tagtggtgtt cgtcgtcggt gagacggcac 1141 gcgccgatca tgtcagcttc aatggctatg agcgcgatac tttcccacag cttgccaaga 1201 tcgatggcgt gaccaatttt agcaatgtca catcgtgcgg cacatcgacg gcgtattctg 1261 tgccgtgtat gttcagctat ctgggcgcgg atgagtatga tgtcgatacc gccaaatacc 1321 aagaaaatgt gctggatacg ctggatcgct tgggcgtaag tatcttgtgg cgtgataata 1381 attcggactc aaaaggcgtg atggataagc tgccaaaagc gcaatttgcc gattataaat 1441 ccgcgaccaa caacgccatc tgcaacacca atccttataa cgaatgccgc gatgtcggta 1501 tgctcgttgg cttagatgac tttgtcgctg ccaataacgg caaagatatg ctgatcatgc 1561 tgcaccaaat gggcaatcac gggcctgcgt attttaagcg atatgatgaa aagtttgcca 1621 aattcacgcc agtgtgtgaa ggtaatgagc ttgccaagtg cgaacatcag tccttgatca 1681 atgcttatga caatgccttg cttgccaccg atgatttcat cgctcaaagt atccagtggc 1741 tgcagacgca cagcaatgcc tatgatgtct caatgctgta tgtcagcgat catggcgaaa 1801 gtctgggtga gaacggtgtc tatctacatg gtatgccaaa tgcctttgca ccaaaagaac 1861 agcgcagtgt gcctgcattt ttctggacgg ataagcaaac tggcatcacg ccaatggcaa 1921 ccgataccgt cctgacccat gacgcgatca cgccgacatt attaaagctg tttgatgtca 1981 ccgcggacaa agtcaaagac cgcaccgcat tcatccgctg actcgagcac caccaccacc 2041 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg 2101 ctgagcaaaa ggccatccgt caggatggcc ttctggtggc acttttcggg gaaatgtgcg 2161 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta 2221 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat 2281 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt 2341 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac 2401 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga 2461 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag 2521 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg 2581 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa 2641 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag 2701 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg 2761 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc 2821 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca 2881 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc 2941 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc 3001 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt 3061 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc 3121 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 3181 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 3241 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 3301 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 3361 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 3421 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 3481 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 3541 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 3601 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 3661 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 3721 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 3781 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 3841 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag 3901 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg 3961 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg 4021 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg 4081 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta 4141 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc 4201 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat 4261 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct 4321 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt 4381 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac 4441 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact 4501 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt 4561 taatacagat gtaggtgttc cacagggt //