LOCUS       Exported                6035 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFS83_PhCMV-LuxS-pA, complete sequence.
ACCESSION   MF688636
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6035)
  AUTHORS   Sedlmayer F, Hell D, Mueller M, Auslaender D, Fussenegger M.
  TITLE     Designer cells programming quorum-sensing interference with human 
            pathogens
  JOURNAL   Nat Commun (2018) In press
REFERENCE   2  (bases 1 to 6035)
  AUTHORS   Sedlmayer F, Hell D, Mueller M, Auslaender D, Fussenegger M.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-AUG-2017) Department of Biosystems Science and 
            Engineering (D-BSSE), ETH Zurich, Mattenstrasse 26, Baseel 4058, 
            Switzerland
REFERENCE   3  (bases 1 to 6035)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6035
                     /organism="Cloning vector pFS83_PhCMV-LuxS-pA"
                     /mol_type="other DNA"
                     /db_xref="taxon:2138157"
     CDS             12..527
                     /codon_start=1
                     /product="S-ribosylhomocysteine lyase"
                     /label=S-ribosylhomocysteine lyase
                     /note="LuxS"
                     /protein_id="AVR59183.1"
                     /translation="MPLLDSFTVDHTRMEAPAVRVAKTMNTPHGDAITVFDLRFCVPNK
                     EVMPERGIHTLEHLFAGFMRNHLNGNGVEIIDISPMGCRTGFYMSLIGTPDEQRVADAW
                     KAAMEDVLKVQDQNQIPELNVYQCGTYQMHSLQEAQDIARSILERDVRINSNEELALPK
                     EKLQELHI"
     regulatory      568..795
                     /regulatory_class="polyA_signal_sequence"
                     /note="bovine growth hormone polyadenylation signal 
                     sequence; bgh-PolyA"
     polyA_signal    571..795
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      841..1269
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1283..1612
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1463..1598
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1661..2686
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    2816..2937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2986..3002)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3010..3026
                     /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(3034..3064)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3079..3100
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3388..3973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4144..5004)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5005..5109)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5375..5754
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      5376..5992
                     /regulatory_class="promoter"
                     /note="human cytomegalovirus immediate early enhancer and 
                     promoter; PhCMV Promoter"
     promoter        5755..5958
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        6003..6021
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"