LOCUS       sequence-219864-        3922 bp    DNA     circular SYN 15-DEC-2025
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    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.chembiol.2016.11.011"; journalName: "Cell Chem Biol"; 
            date: "2017-01-19- 19"; volume: "24"; issue: "1"; pages: "98-109"
COMMENT     Sequence Label: pGDP1 NDM-1
FEATURES             Location/Qualifiers
     source          1..3922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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      complement(1259..1286)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(1377..2192)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     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
//
