LOCUS       Exported                6812 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gene replacement vector pLAW344, complete sequence.
ACCESSION   DQ289040
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6812)
  AUTHORS   Edelstein PH, Hu B.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-NOV-2005) Pathology & Lab Medicine, Univ of 
            Pennsylvania School of Medicine, Clinical Microbiology 4 Gates 3400 
            Spruce St, Philadelphia, PA 19104-4283, USA
REFERENCE   2  (bases 1 to 6812)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6812
                     /organism="Gene replacement vector pLAW344"
                     /lab_host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /note="used as a gene replacement vector in Legionella 
                     pneumophila"
                     /db_xref="taxon:359910"
     CDS             complement(205..1065)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /note="ampicillin resistance; Amp-R"
                     /protein_id="ABC02197.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(205..1065)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(1066..1170)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    1301..1334
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(1301..1334)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            complement(1368..3275)
                     /gene="sacB"
                     /label=sacB
     CDS             complement(1391..2812)
                     /codon_start=1
                     /gene="sacB"
                     /product="SacB"
                     /label=sacB
                     /note="sucrase"
                     /protein_id="ABC02199.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        complement(2813..3258)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     primer_bind     complement(3333..3349)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            3720..3831
                     /label=RK2
                     /note="RK2"
     oriT            3722..3831
                     /note="incP origin of transfer"
     CDS             3864..4235
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     promoter        4705..4807
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4808..5467
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="degrades chloramphenicol"
                     /protein_id="ABC02198.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            4808..5467
                     /gene="cat"
                     /label=cat
     misc_feature    5743..5776
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(5743..5776)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    5932..6072
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(6258..34)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      6258..6812
                     /label=oriR (ColE 1)
                     /note="oriR (ColE 1)"