LOCUS       Exported                5846 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pEX100T, complete sequence.
ACCESSION   U17500
VERSION     .
KEYWORDS    beta-lactamase (bla); lac alpha peptide (lacZalpha); levansucr...
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5846)
  AUTHORS   Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-NOV-1994) Herbert P. Schweizer, Microbiology & 
            Infectious Diseases, University of Calgary, 3330 Hospital Drive 
            N.W., Calgary, Alberta T2N 4N1, Canada
REFERENCE   2  (bases 1 to 5846)
  AUTHORS   Schweizer HP, Hoang TT.
  TITLE     An improved system for gene replacement and xylE fusion analysis in 
            Pseudomonas aeruginosa
  JOURNAL   Gene 158 (1), 15-22 (1995)
  PUBMED    7789804
REFERENCE   3  (bases 1 to 5846)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5846
                     /organism="Cloning vector pEX100T"
                     /host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:37970"
     CDS             complement(209..1630)
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /protein_id="AAA57325.1"
                     /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"
     misc_feature    2104..2928
                     /label=region containing oriT
                     /note="region containing oriT"
     oriT            2450..2559
                     /note="incP origin of transfer"
     misc_feature    complement(2984..3309)
                     /label=E. coli 5S RNA and terminators
                     /note="E. coli 5S RNA and terminators"
     regulatory      complement(2984..3011)
                     /regulatory_class="terminator"
                     /note="T2"
     terminator      2984..3011
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      complement(3018..3143)
                     /regulatory_class="terminator"
                     /note="T1"
     rRNA            complement(3022..3309)
                     /product="5S ribosomal RNA"
     terminator      3103..3189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3345..3629)
                     /codon_start=1
                     /gene="lacZalpha"
                     /product="lac alpha peptide"
                     /label=lacZalpha
                     /protein_id="AAA57324.1"
                     /translation="MTMITPSSRDNRVIPGRDNRVMSLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRAWLFWRMREDFQPDTD"
     gene            complement(3345..3629)
                     /gene="lacZalpha"
                     /label=lacZalpha
     primer_bind     3539..3555
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3565..3605
                     /label=polylinker
                     /note="polylinker"
     primer_bind     complement(3625..3641)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3649..3665
                     /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(3673..3703)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3718..3739
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4027..4615)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4786..5646)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAA57323.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4786..5646)
                     /gene="bla"
                     /label=bla
     promoter        complement(5647..5751)
                     /gene="bla"
                     /label=AmpR promoter