LOCUS       Exported                4591 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGDP2, complete sequence.
ACCESSION   KX859085
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4591)
  AUTHORS   Stokes JM, MacNair CR, Ilyas B, French S, Cote J-P., Farha MA, 
            Sieron AO, Coombes BK, Brown ED.
  TITLE     An antiprotozoal drug overcomes acquired colistin resistance
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4591)
  AUTHORS   Stokes JM, MacNair CR, Ilyas B, French S, Cote J-P., Farha MA, 
            Sieron AO, Coombes BK, Brown ED.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-SEP-2016) Biochemistry and Biomedical Sciences, 
            McMaster University, 1200 Main Street West, Hamilton, Ontario L8N 
            3Z5, Canada
REFERENCE   3  (bases 1 to 4591)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4591
                     /organism="Cloning vector pGDP2"
                     /lab_host="Escherichia coli strain BW25113"
                     /mol_type="other DNA"
                     /db_xref="taxon:1961793"
     RBS             150..172
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             179..1804
                     /codon_start=1
                     /gene="mcr-1"
                     /product="Mcr-1"
                     /EC_number="2.7.4.30"
                     /label=mcr-1
                     /note="Mcr-1 phosphoethanolamine transferase"
                     /protein_id="AQS99092.1"
                     /translation="MVQHTSVWYRRSVSPFVLVASVAVFLTATANLTFFDKISQTYPIA
                     DNLGFVLTIAVVLFGAMLLITTLLSSYRYVLKPVLILLLIMGAVTSYFTDTYGTVYDTT
                     MLQNALQTDQAETKDLLNAAFIMRIIGLGVLPSLLVAFVKVDYPTWGKGLMRRLGLIVA
                     SLALILLPVVAFSSHMASFFRVHKPLRSYVNPIMPIYSVGKLASIEYKKASAPKDTIYH
                     AKDAVQATKPDMRKPRLVVFVVGETARADHVSFNGYERDTFPQLAKIDGVTNFSNVTSC
                     GTSTAYSVPCMFSYLGADEYDVDTAKYQENVLDTLDRLGVSILWRDNNSDSKGVMDKLP
                     KAQFADYKSATNNAICNTNPYNECRDVGMLVGLDDFVAANNGKDMLIMLHQMGNHGPAY
                     FKRYDEKFAKFTPVCEGNELAKCEHQSLINAYDNALLATDDFIAQSIQWLQTHSNAYDV
                     SMLYVSDHGESLGENGVYLHGMPNAFAPKEQRSVPAFFWTDKQTGITPMATDTVLTHDA
                     ITPTLLKLFDVTADKVKDRTAFIR"
     gene            179..1804
                     /gene="mcr-1"
                     /label=mcr-1
     CDS             1811..1828
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1890..1917
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(2008..2823)
                     /codon_start=1
                     /gene="kanR"
                     /product="KanR"
                     /EC_number="2.7.1.95"
                     /label=kanR
                     /note="amino glycoside phosphotransferase"
                     /protein_id="AQS99093.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(2008..2823)
                     /gene="kanR"
                     /label=kanR
     rep_origin      2921..3551
     rep_origin      2945..3533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3719..3861
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3963..4154)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"