LOCUS       sequence-219874-        4036 bp    DNA     circular SYN 15-APR-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 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 Apr 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: pGDP3 aph(3'')-Ia
FEATURES             Location/Qualifiers
     source          1..4036
                     /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             1161..1178
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      complement(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             join(1397..1465,1466..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
//
