LOCUS       Exported                6921 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pKKM801 DNA, complete sequence.
ACCESSION   LC331101
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6921)
  AUTHORS   Sakka K, Wang YY, Sakka M, Kunitake E, Kimura T.
  TITLE     A plasmid vector for Ruminiclostridium josui
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6921)
  AUTHORS   Sakka K, Kimura T.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-OCT-2017) Contact:Kazuo Sakka Mie University, Graduate
            School of Bioresources; 1577 Kurimamachiya-cho, Tsu, Mie 514-8507, 
            Japan URL :http://www.bio.mie-u.ac.jp/
REFERENCE   3  (bases 1 to 6921)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6921
                     /organism="Expression vector pKKM801"
                     /mol_type="other DNA"
                     /db_xref="taxon:2049427"
     promoter        90..194
                     /gene="bla"
                     /label=AmpR promoter
     CDS             195..1055
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="BBA94092.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            195..1055
                     /gene="bla"
                     /label=bla
     rep_origin      1215..1829
                     /label=putative oriEC
                     /note="putative oriEC"
     rep_origin      1226..1814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2102..2123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2138..2168
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2176..2192
                     /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     2200..2216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2232..2275
                     /regulatory_class="promoter"
                     /note="pcip promoter"
     RBS             2558..2566
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     misc_feature    2766..2844
                     /label=multiple cloning site and His-tag
                     /note="multiple cloning site and His-tag"
     CDS             2824..2841
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2854..2870)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3458..3687
                     /note="oriCP; C. perfringens pIP404 replication origin"
     CDS             complement(3750..4970)
                     /codon_start=1
                     /gene="rep"
                     /product="pIP404 replication"
                     /label=rep
                     /protein_id="BBA94093.1"
                     /translation="MRNALKLNKKNYIDNIHSRSKGWITRSVIDKKGYSQWHYKYAELK
                     DLDMSDENIYITLNTFYKPCRRLENIKELNTLFIDLDYYKTGKTKDQVLMDLEKNYFNQ
                     SIPIPNYVIDSGRGMYLIWIINAVPSKALPLWKAVQEYLYNQLKYFGADRQALDATRIL
                     RVPGSINSKSKTVVNILDEYEYIYDLREIQNGFLPELKPYERKKGRPSKINYIYRERSL
                     YYGRIQDIIKLCELREYDLKGHRELILFLYRYYLCSFTEDIEKALNDVLELNSMFRQHL
                     SEREVIRATRSAERCYLDKNKQYKYKNETLIELLEITEEEQKYMTIIISKKEYKRRENI
                     RGKKNYQEQLKAKGKATKKEELNVLRKKIKALKEKGFKNKEITLMLEVPIKTLERHITY
                     MKKNGLL"
     gene            complement(3750..4970)
                     /gene="rep"
                     /label=rep
     CDS             complement(6029..6766)
                     /codon_start=1
                     /gene="ermBP"
                     /product="RNA methylase"
                     /label=ermBP
                     /protein_id="BBA94094.1"
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
                     KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIPYH
                     LSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
                     PKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHAKVNN
                     LSTVTYEQVLSIFNSYLLFNGRK"
     gene            complement(6029..6766)
                     /gene="ermBP"
                     /label=ermBP