LOCUS       Exported                4849 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMM23, complete sequence.
ACCESSION   MH697727
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4849)
  AUTHORS   Dean C, Barkan D, Bermingham A, Blais J, Casey F, Casarez A, Colvin 
            R, Fuller J, Jones A, Li C, Lopez S, Metzger L, Prathapam R, Rasper 
            D, Reck F, Ruzin A, Shaul J, Shen X, Simmons R, Skewes-Cox P, 
            Tamrakar P, Uehara T.
  TITLE     The monobactam LYS228: mode of action and mechanisms decreasing in 
            vitro susceptibility of Escherichia coli and Klebsiella pneumoniae
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4849)
  AUTHORS   Wei J-R., Mostafavi M, Takeoka K.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-JUL-2018) Infectious Diseases, Bacteriology, Novartis 
            Institutes for Biomedical Research, 5300 Chiron way, Emeryville, CA 
            94608, USA
REFERENCE   3  (bases 1 to 4849)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4849
                     /organism="Cloning vector pMM23"
                     /mol_type="other DNA"
                     /db_xref="taxon:2303769"
     CDS             complement(80..1501)
                     /codon_start=1
                     /gene="sacB"
                     /product="SacB"
                     /label=sacB
                     /protein_id="AXP32211.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(80..1501)
                     /gene="sacB"
                     /label=sacB
     promoter        1738..1766
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1955..2488
                     /codon_start=1
                     /gene="accC1"
                     /product="aminoglycoside 3-N-acetyltransferase"
                     /label=accC1
                     /protein_id="AXP32212.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            1955..2488
                     /gene="accC1"
                     /label=accC1
     promoter        2794..2822
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     misc_RNA        2828..2847
                     /product="gRNA variable region"
     misc_RNA        2848..2923
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      2945..2979
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     rep_origin      3019..3474
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      3585..4173
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4359..4499
                     /label=bom
                     /note="basis of mobility region from pBR322"