LOCUS       Exported                5831 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pEX18Gm, complete sequence.
ACCESSION   AF047518
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5831)
  AUTHORS   Hoang TT, Karkhoff-Schweizer RR, Kutchma AJ, Schweizer HP.
  TITLE     A broad-host-range Flp-FRT recombination system for site-specific 
            excision of chromosomally-located DNA sequences: application for 
            isolation of unmarked Pseudomonas aeruginosa mutants
  JOURNAL   Gene 212 (1), 77-86 (1998)
  PUBMED    9661666
REFERENCE   2  (bases 1 to 5831)
  AUTHORS   Hoang TT, Karkhoff-Schweizer RR, Kutchma AJ, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-FEB-1998) Microbiology, Colorado State University, 
            Fort Collins, CO 80523, USA
REFERENCE   3  (bases 1 to 5831)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5831
                     /organism="Cloning vector pEX18Gm"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:73159"
     CDS             complement(209..1630)
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase precursor"
                     /label=sacB
                     /protein_id="AAC27023.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(209..1630)
                     /gene="sacB"
                     /label=sacB
     promoter        complement(1631..2076)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     oriT            2104..2928
     oriT            2450..2559
                     /note="incP origin of transfer"
     terminator      2980..3007
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      complement(2980..3006)
                     /regulatory_class="terminator"
                     /note="T2"
     terminator      3099..3185
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      complement(3139..3182)
                     /regulatory_class="terminator"
                     /note="T1"
     rRNA            complement(3186..3305)
                     /product="Eschericia coli 5S ribosomal RNA"
     CDS             complement(3341..3625)
                     /codon_start=1
                     /gene="lacZalpha"
                     /product="lac alpha peptide"
                     /label=lacZalpha
                     /protein_id="AAC27024.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRAWLFWRMREDFQPDTD"
     gene            complement(3341..3625)
                     /gene="lacZalpha"
                     /label=lacZalpha
     primer_bind     3535..3551
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3555..3611
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3621..3637)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3645..3661
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3669..3699)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3714..3735
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4023..4611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4864..5397)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamicin acetyl transferase"
                     /label=aacC1
                     /protein_id="AAC27025.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(4864..5397)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(5586..5614)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(5632..5736)
                     /gene="bla"
                     /label=AmpR promoter