LOCUS       Exported                7235 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pME8, complete sequence.
ACCESSION   KC679682
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7235)
  AUTHORS   Audtho M, Ratlertkarn M, Wiwat C.
  TITLE     pME8, an improved shuttle vector for expression in Bacillus spp. and
            E. coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7235)
  AUTHORS   Audtho M, Ratlertkarn M, Wiwat C.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-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 7235)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7235
                     /organism="Shuttle vector pME8"
                     /mol_type="other DNA"
                     /label=derived from Bacillus cereus sequence
                     /note="derived from Bacillus cereus sequence"
                     /db_xref="taxon:1296590"
     misc_feature    7..105
                     /note="contains cloning sites SmaI, PstI, EcoRI, EcoRV, 
                     HindIII, BglII, SalI, NotI, SacI, SacII, PstI, XbaI, XhoI, 
                     ApaI, and KpnI"
     promoter        complement(118..136)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(153..171)
                     /label=M13 reverse primer
                     /note="M13 reverse primer"
     primer_bind     complement(157..173)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(172..191)
                     /label=M13 pUC reverse primer
                     /note="M13 pUC reverse primer"
     protein_bind    181..197
                     /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(202..235)
                     /regulatory_class="promoter"
                     /note="from lac gene"
     promoter        complement(205..235)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    250..271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(559..1147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1318..2178)
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="AGH24528.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2179..2283)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(2220..2246)
                     /regulatory_class="promoter"
                     /note="from ampicillin resistance protein gene"
     rep_origin      2310..2765
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      2327..2633
                     /label=f1
                     /note="f1"
     primer_bind     2891..2913
                     /label=M13 forward primer
                     /note="M13 forward primer"
     primer_bind     2906..2922
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2929..2947
                     /label=T7 promoter primer
                     /note="T7 promoter primer"
     promoter        2929..2947
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_difference 3389
                     /replace="g"
                     /label=G > C to delete EcoRI site in pBC16
                     /note="G > C to delete EcoRI site in pBC16"
     CDS             3807..4811
                     /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             3828..4811
                     /codon_start=1
                     /gene="repU"
                     /product="RepU"
                     /label=repU
                     /protein_id="AGH24529.1"
                     /translation="MCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDYFELLHIL
                     EFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGIQSQKVVA
                     EVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLVGFMRATE
                     VTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPNVKVQMIR
                     PKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLISYGGLLKE
                     IHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            3828..4811
                     /gene="repU"
                     /label=repU
     CDS             5011..6387
                     /codon_start=1
                     /product="tetracycline resistance protein"
                     /label=tetracycline resistance protein
                     /protein_id="AGH24531.1"
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     misc_difference 5011
                     /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 6445
                     /replace="g"
                     /note="G > C to delete EcoRI site in DNA fragment from 
                     pBC16"
     rep_origin      6504..6780
                     /label=ori L
                     /note="ori L"
     CDS             6951..234
                     /codon_start=1
                     /gene="mob"
                     /product="Mob"
                     /label=mob
                     /note="required for plasmid mobilization"
                     /protein_id="AGH24530.1"
                     /translation="MSYAVCRMQKVKSAGLKGMQFHNQRERKSRTNDDIDHERTRENYD
                     LKNDKNIDYNERVKEIIESQKTGTRKTRKDAVLVNELLVTSDRDFFEQLDRSPGLQEFD
                     IKLRSTVDAAASSRGCSLDLEGGPGTQLLFPLVRVNFELGVIMVIAVSCVKLLSAHNST
                     QHTSRKHKV"
     gene            6951..234
                     /gene="mob"
                     /label=mob