LOCUS       Exported                6168 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gene deletion vector pEX100Tlink, complete sequence.
ACCESSION   AY906830
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6168)
  AUTHORS   Quenee L, Lamotte D, Polack B.
  TITLE     Combined sacB-based negative selection and cre-lox antibiotic marker
            recycling for efficient gene deletion in pseudomonas aeruginosa
  JOURNAL   BioTechniques 38 (1), 63-67 (2005)
  PUBMED    15679087
REFERENCE   2  (bases 1 to 6168)
  AUTHORS   Quenee L, Lamotte D, Polack B.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-JAN-2005) GREPI, CHU, Hopital Michallon, Grenoble 
            38043, France
REFERENCE   3  (bases 1 to 6168)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6168
                     /organism="Gene deletion vector pEX100Tlink"
                     /lab_host="Pseudomonas aeruginosa"
                     /mol_type="other DNA"
                     /label=derived from Cloning vector pEX100T
                     /note="derived from Cloning vector pEX100T"
                     /db_xref="taxon:313915"
     CDS             complement(209..1630)
                     /codon_start=1
                     /gene="sacB"
                     /product="SacB"
                     /label=sacB
                     /note="levansucrase"
                     /protein_id="AAX11247.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(209..1630)
                     /gene="sacB"
                     /label=sacB
     mRNA            complement(1631..208)
     promoter        complement(1631..2076)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     misc_feature    2104..2928
                     /label=contains oriT
                     /note="contains oriT"
     oriT            2450..2559
                     /note="incP origin of transfer"
     regulatory      complement(2984..3011)
                     /regulatory_class="terminator"
     terminator      2984..3011
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      complement(3018..3143)
                     /regulatory_class="terminator"
     rRNA            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..3746)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFARGRDNRVMSLALAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDR
                     PSQQLRSLNGEWRAWLFWRMREDFQPDTD"
     primer_bind     3539..3555
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3656..3672
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3676..3732
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3742..3758)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3766..3782
                     /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(3790..3820)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3835..3856
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3947..3963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3971..3987
                     /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(3995..4025)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4040..4061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4349..4937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5108..5968)
                     /codon_start=1
                     /gene="bla"
                     /product="bla"
                     /label=bla
                     /note="beta-lactamase"
                     /protein_id="AAX11248.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5108..5968)
                     /gene="bla"
                     /label=bla
     mRNA            complement(5969..5107)
     promoter        complement(5969..6073)
                     /gene="bla"
                     /label=AmpR promoter