LOCUS       Exported                7485 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pME9, complete sequence.
ACCESSION   KC679683
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7485)
  AUTHORS   Audtho M, Ratlertkarn M, Promdonkoy B.
  TITLE     pME9, a new vector for sporulation stage expression in Bacillus spp.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7485)
  AUTHORS   Audtho M, Ratlertkarn M, Promdonkoy B.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-FEB-2013) National Center for Genetic Engineering and 
            Biotechnology, National Science and Technology Development Agency, 
            113 Phaholyothin Road, Klong Nueng, Klong Luang, Pathum Thani 12120,
            Thailand
REFERENCE   3  (bases 1 to 7485)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7485
                     /organism="Expression vector pME9"
                     /mol_type="other DNA"
                     /label=derived from Bacillus cereus sequence
                     /note="derived from Bacillus cereus sequence"
                     /db_xref="taxon:1296589"
     misc_feature    13..18
                     /label=CTGCAG recognition site for PstI
                     /note="CTGCAG recognition site for PstI"
     regulatory      18..270
                     /regulatory_class="promoter"
                     /note="from Bacillus thuringiensis cyt2Aa gene"
     misc_feature    269..355
                     /note="contains cloning sites EcoRI, EcoRV, HindIII, BglII,
                     SalI, NotI, SacI, SacII, PstI, XbaI, XhoI, ApaI, KpnI"
     promoter        complement(368..386)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(403..421)
                     /label=M13 reverse primer
                     /note="M13 reverse primer"
     primer_bind     complement(407..423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(422..441)
                     /label=M13 pUC reverse primer
                     /note="M13 pUC reverse primer"
     protein_bind    431..447
                     /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)."
     regulatory      complement(452..485)
                     /regulatory_class="promoter"
                     /note="from lac gene"
     promoter        complement(455..485)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    500..521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(809..1397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1568..2428)
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="AGH24532.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2429..2533)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(2470..2796)
                     /regulatory_class="promoter"
                     /note="from ampicillin resistance protein gene"
     rep_origin      2560..3015
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      2577..2883
                     /label=f1
                     /note="f1"
     primer_bind     3141..3153
                     /label=M13 forward primer
                     /note="M13 forward primer"
     primer_bind     3156..3172
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3179..3197
                     /label=T7 promoter primer
                     /note="T7 promoter primer"
     promoter        3179..3197
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_difference 3539
                     /replace="g"
                     /note="G > C to delete EcoRI site of DNA fragment from 
                     pBC16"
     CDS             4057..5061
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     CDS             4078..5061
                     /codon_start=1
                     /gene="repU"
                     /product="RepU"
                     /label=repU
                     /protein_id="AGH24533.1"
                     /translation="MCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDYFELLHIL
                     EFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGIQSQKVVA
                     EVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLVGFMRATE
                     VTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPNVKVQMIR
                     PKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLISYGGLLKE
                     IHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            4078..5061
                     /gene="repU"
                     /label=repU
     CDS             5261..6637
                     /codon_start=1
                     /product="tetracycline resistance protein"
                     /label=tetracycline resistance protein
                     /note="for selection in tetracycline containing media"
                     /protein_id="AGH24534.1"
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     misc_difference 5261
                     /replace="g"
                     /note="G > A to change start codon from GTG to ATG for 
                     improvement of transformation efficiency in the media 
                     containing tetracycline"
     misc_difference 6695
                     /replace="g"
                     /note="G > C to delete EcoRI site in DNA fragment from 
                     pBC16"
     rep_origin      6754..7030
                     /label=ori L
                     /note="ori L"
     misc_feature    7201..7485
                     /label=from mob gene
                     /note="from mob gene"