LOCUS       Exported                3101 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 3101)
  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 3101)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3101
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             438..629
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(623..645)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    731..873
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     786..805
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(888..905)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1059..1647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1139..1158)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1758..1777)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1769..2584
                     /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     complement(1842..1861)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2675..2702
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(2764..2781)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2831..2863)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(2867..2884)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(2894..2911)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2930..2952
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 agatctcccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac
       61 gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa
      121 tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca
      181 ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg
      241 tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata
      301 ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc
      361 gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc
      421 cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta
      481 tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg
      541 cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc
      601 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
      661 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
      721 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
      781 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atatgcggtg tgaaataccg
      841 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
      901 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
      961 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1021 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1081 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1141 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1201 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1261 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1321 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1381 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1441 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     1501 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1561 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1621 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgaacaataa aactgtctgc
     1741 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc
     1801 taggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1861 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
     1921 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     1981 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2041 tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga
     2101 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2161 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2221 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2281 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2341 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2401 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2461 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     2521 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2581 ctaagaatta attcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2641 gggttccgcg cacatttccc cgaaaagtgc caccagaagg ccatcctgac ggatggcctt
     2701 ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg gctttgttag cagccggatc
     2761 tcagtggtgg tggtggtggt gctcgagtgc ggccgcaagc ttgtcgacgg agctcgaatt
     2821 cggatccgcg acccatttgc tgtccaccag tcatgctagc catatggctg ccgcgcggca
     2881 ccaggccgct gctgtgatga tgatgatgat ggctgctgcc catggtatat ctccttctta
     2941 aagttaaaca aaattatttc tagaccgggc gctatcatgc cataccgcga aaggttttgc
     3001 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     3061 cagcccagta gtaggttgag gccgttgagc accgccgccg c
//
