LOCUS       Exported                6714 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLMF04, complete sequence.
ACCESSION   KU997671
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6714)
  AUTHORS   Liu M, Wang M.
  TITLE     Development of a plasmid-based methods for expression and antisense 
            gene knockdown in Riemerella anatipestifer and application
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6714)
  AUTHORS   Liu M, Wang M.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-MAR-2016) Institute of Preventive Veterinary Medicine,
            Sichuan Agricultural University, No. 211, Huimin Road, Chengdu, 
            Sichuan 611130, China
REFERENCE   3  (bases 1 to 6714)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6714
                     /organism="Cloning vector pLMF04"
                     /mol_type="other DNA"
                     /label=shuttle vector
                     /note="shuttle vector"
                     /db_xref="taxon:1882323"
     protein_bind    290..311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        326..356
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    364..380
                     /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)."
     primer_bind     388..404
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2671..2688
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3357..4322)
                     /codon_start=1
                     /product="beta-lactamase CfxA"
                     /function="cefoxitin resistance protein"
                     /label=beta-lactamase CfxA
                     /protein_id="ANW35657.1"
                     /translation="MEKNRKKQIVVLSIALVCIFILVFSLFHKSATKDSANPPLTNVLT
                     DSISQIVSACPGEIGVAVIVNNRDTVKVNNKSVYPMMSVFKVHQALALCNDFDNKGISL
                     DTLVNINRDKLDPKTWSPMLKDYSGPVISLTVRDLLRYTLTQSDNNASNLMFKDMVNVA
                     QTDSFIATLIPRSSFQIAYTEEEMSADHNKAYSNYTSPLGAAMLMNRLFTEGLIDDEKQ
                     SFIKNTLKECKTGVDRIAAPLLDKEGVVIAHKTGSGYVNENGVLAAHNDVAYICLPNNI
                     SYTLAVFVKDFKGNKSQASQYVAHISAVVYSLLMQTSVKS"
     primer_bind     complement(4502..4518)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4992..5096
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5097..5957
                     /codon_start=1
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=beta-lactamase
                     /protein_id="ANW35656.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6128..2
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"