pGDP2 MCR-1

LOCUS       pGDP2 MCR-1               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
//