LOCUS       Exported                8358 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWW3323, complete sequence.
ACCESSION   KY776506
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8358)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Tunable expression tools enable single-cell strain distinction in 
            the gut microbiome
  JOURNAL   Cell (2017) In press
REFERENCE   2  (bases 1 to 8358)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-MAR-2017) Microbiology & Immunology, Stanford 
            University School of Medicine, 299 Campus Drive, Stanford, CA 
            94305-5124, USA
REFERENCE   3  (bases 1 to 8358)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8358
                     /organism="Cloning vector pWW3323"
                     /mol_type="other DNA"
                     /db_xref="taxon:1977332"
     rep_origin      25..798
     rep_origin      29..417
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            588..697
                     /note="incP origin of transfer"
     regulatory      841..933
                     /regulatory_class="promoter"
     regulatory      942..964
                     /regulatory_class="ribosome_binding_site"
     CDS             965..2467
                     /codon_start=1
                     /product="Bxb1"
                     /label=Bxb1
                     /protein_id="ARI71322.1"
                     /translation="MRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLD
                     VSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLV
                     VSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGY
                     LPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQG
                     REPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAE
                     LVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTV
                     AMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAG
                     SPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSM
                     NVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     terminator      2487..2514
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      2489..2518
                     /regulatory_class="terminator"
     CDS             complement(2746..4029)
                     /codon_start=1
                     /product="NBU integrase"
                     /label=NBU integrase
                     /protein_id="ARI71323.1"
                     /translation="MNIKRNIIFALESRKKNGVPIVENVPIRMRVIFASQRIEFTTGYR
                     IDVAKWDADKQRVKSGCTNKLKQSAAEINTDLLKYYAEIQNIFKEFEVQEVMPTTQQLK
                     EAFNMRMKDTSEEQPEEAPVSFWEVFDEFVKECGNQNNWTASTYEKFAAVRNHLKEFKE
                     DATFNYFNEFGLNEYVNFLRDTKDMRNSTIGKQMGFLKWFLRWSFKKGHHQNIAYDTFK
                     PKLKTTSKKVIFLTWDELNKLKDYQIPKDKQYLERVRDVFLFCCFTSLRYSDVRNLKRS
                     DVKSDHIEITTVKTADSLTIELNKYSKAILDKYKDIHFENYMALPVISNQKMNDYLKEL
                     GELAEINEPVRETYYKGNERIDEVTPKYALLSTHAGRRTFICNALALGIPAQVVMKWTG
                     HSDYKAMKPYIDIADDIKANAMNKFNQL"
     misc_feature    4189..4200
                     /label=integration site
                     /note="integration site"
     misc_feature    4474..5071
                     /label=split antibiotic resistance marker
                     /note="split antibiotic resistance marker"
     CDS             complement(4475..5335)
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ARI71324.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    5068..5335
                     /label=split antibiotic resistance marker
                     /note="split antibiotic resistance marker"
     promoter        complement(5336..5440)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(5868..7793)
                     /codon_start=1
                     /product="TetQ"
                     /label=TetQ
                     /protein_id="ARI71321.1"
                     /translation="MNIINLGILAHIDAGKTSVTENLLFASGATEKCGCVDNGDTITDS
                     MDIEKRRGITVRASTTSIIWNGVKCNIIDTPGHMDFIAEVERTFKMLDGAVLILSAKEG
                     IQAQTKLLFNTLQKLQIPTIIFINKIDRAGVNLERLYLDIKANLSQDVLFMQNVVDGSV
                     YPVCSQTYIKEEYKEFVCNHDDNILERYLADSEISPADYWNTIIALVAKAKVYPVLHGS
                     AMFNIGINELLDAITSFILPPASVSNRLSSYLYKIEHDPKGHKRSFLKIIDGSLRLRDV
                     VRINDSEKFIKIKNLKTINQGREINVDEVGANDIAIVEDMDDFRIGNYLGAEPCLIQGL
                     SHQHPALKSSVRPDRPEERSKVISALNTLWIEDPSLSFSINSYSDELEISLYGLTQKEI
                     IQTLLEERFSVKVHFDEIKTIYKERPVKKVNKIIQIEVPPNPYWATIGLTLEPLPLGTG
                     LQIESDISYGYLNHSFQNAVFEGIRMSCQSGLHGWEVTDLKVTFTQAEYYSPVSTPADF
                     RQLTPYVFRLALQQSGVDILEPMLYFELQIPQAASSKAITDLQKMMSEIEDISCNNEWC
                     HIKGKVPLNTSKDYASEVSSYTKGLGIFMVKPCGYQITKGGYSDNIRMNEKDKLLFMFQ
                     KSMSSK"