LOCUS       Exported                7850 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pME10, complete sequence.
ACCESSION   KT350984
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7850)
  AUTHORS   Audtho M, Maureen K, Ratlertkarn M, Rattanalapho C.
  TITLE     pME13, an improved expression vector for vegetative stage of 
            Bacillus spp.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7850)
  AUTHORS   Audtho M, Maureen K, Ratlertkarn M, Rattanalapho C.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUL-2015) 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 7850)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7850
                     /organism="Cloning vector pME10"
                     /mol_type="other DNA"
                     /db_xref="taxon:1770536"
     regulatory      25..652
                     /regulatory_class="promoter"
                     /note="from Bacillus thuringiensis subsp. tenebrionis"
     misc_feature    652..719
                     /note="multiple cloning site; 
                     BglII-SalI-NotI-SacI-SacII-PstI-XbaI-XhoI-ApaI-KpnI"
     promoter        complement(733..751)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(768..786)
                     /label=M13 reverse
                     /note="M13 reverse"
     primer_bind     complement(772..788)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(787..806)
                     /label=M13 pUC reverse
                     /note="M13 pUC reverse"
     protein_bind    796..812
                     /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(817..850)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(820..850)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    865..886
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1174..1762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1933..2793)
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="ALT14585.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2794..2898)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2925..3380
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      2942..3248
                     /label=f1 origin
                     /note="f1 origin"
     primer_bind     3506..3528
                     /label=M13 forward
                     /note="M13 forward"
     primer_bind     3521..3537
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3544..3562
                     /label=T7
                     /note="T7"
     promoter        3544..3562
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3571..4198
                     /label=mob fragment
                     /note="mob fragment"
     CDS             4422..5426
                     /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             4443..5426
                     /codon_start=1
                     /gene="repU"
                     /product="RepU"
                     /label=repU
                     /note="required for plasmid replication in Bacillus"
                     /protein_id="ALT14587.1"
                     /translation="MCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDYFELLHIL
                     EFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGIQSQKVVA
                     EVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLVGFMRATE
                     VTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPNVKVQMIR
                     PKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLISYGGLLKE
                     IHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            4443..5426
                     /gene="repU"
                     /label=repU
     CDS             5626..7002
                     /codon_start=1
                     /product="tetracycline resistance protein"
                     /label=tetracycline resistance protein
                     /protein_id="ALT14586.1"
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     rep_origin      7119..7395
                     /label=ori L
                     /note="ori L"
     misc_feature    7566..7850
                     /label=mob fragment
                     /note="mob fragment"