LOCUS       Exported                9064 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLW18, complete sequence.
ACCESSION   KC820641
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9064)
  AUTHORS   Delaney NF, Kaczmarek ME, Ward LM, Swanson PK, Lee MC, Marx CJ.
  TITLE     Development of an Optimized Medium, Strain and High-Throughput 
            Culturing Methods for Methylobacterium extorquens
  JOURNAL   PLoS ONE 8 (4), E62957 (2013)
  PUBMED    23646164
REFERENCE   2  (bases 1 to 9064)
  AUTHORS   Delaney NF.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-MAR-2013) Organismic and Evolutionary Biology, Harvard
            University, 16 Divinity Ave., Cambridge, MA 02138, USA
REFERENCE   3  (bases 1 to 9064)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9064
                     /organism="Cloning vector pLW18"
                     /mol_type="other DNA"
                     /db_xref="taxon:1331852"
     CDS             complement(1..861)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin-resistance"
                     /protein_id="AGL45405.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(1..861)
                     /gene="bla"
                     /label=bla
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    997..1548
                     /label=region upstream of celA gene
                     /note="region upstream of celA gene"
     misc_feature    1549..2075
                     /label=region downstream of celC gene
                     /note="region downstream of celC gene"
     CDS             complement(2676..3047)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            3080..3189
                     /note="incP origin of transfer"
     promoter        3826..3854
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             3902..5092
                     /codon_start=1
                     /gene="tet"
                     /product="Tet"
                     /label=tet
                     /note="tetracycline efflux pump"
                     /protein_id="AGL45406.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            3902..5092
                     /gene="tet"
                     /label=tet
     CDS             complement(5270..5929)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="AGL45407.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(5270..5929)
                     /gene="cat"
                     /label=cat
     CDS             complement(6027..7448)
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /note="imparts sucrose sensitivity"
                     /protein_id="AGL45408.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(6027..7448)
                     /gene="sacB"
                     /label=sacB
     promoter        complement(7449..7894)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     rep_origin      complement(8306..8894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"