LOCUS       Exported               12485 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGETS109 DNA, complete sequence.
ACCESSION   AB117035
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12485)
  AUTHORS   Tsuge K, Matsui K, Itaya M.
  TITLE     One step assembly of multiple DNA fragments with a designed order 
            and orientation in Bacillus subtilis plasmid
  JOURNAL   Nucleic Acids Res. 31 (21), E133 (2003)
  PUBMED    14576333
REFERENCE   2  (bases 1 to 12485)
  AUTHORS   Itaya M, Tsuge K, Matsui K.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-AUG-2003) Mitsuhiro Itaya, Mitsubishi Kagaku Institute
            of Life Sciences; 11 Minamiooya, Machida, Tokyo 194-8511, Japan 
            (E-mail:ita@libra.ls.m-kagaku.co.jp, Tel:81-42-724-6254, 
            Fax:81-42-724-6317)
REFERENCE   3  (bases 1 to 12485)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12485
                     /organism="Cloning vector pGETS109"
                     /mol_type="other DNA"
                     /label=Bacillus subtilis-Escherichia coli shuttle vector
                     /note="Bacillus subtilis-Escherichia coli shuttle vector"
                     /db_xref="taxon:242382"
     promoter        10..38
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             86..1276
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline resistance protein"
                     /label=tet
                     /protein_id="BAD00172.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            86..1276
                     /gene="tet"
                     /label=tet
     gene            2175..4143
                     /gene="tetL"
                     /label=tetL
     regulatory      2175..2202
                     /regulatory_class="promoter"
                     /gene="tetL"
                     /standard_name="bsr promoter"
     regulatory      2175..2180
                     /regulatory_class="minus_35_signal"
                     /gene="tetL"
                     /standard_name="bsr promoter"
     regulatory      2197..2202
                     /regulatory_class="minus_10_signal"
                     /gene="tetL"
                     /standard_name="bsr promoter"
     regulatory      2481..2485
                     /regulatory_class="ribosome_binding_site"
                     /gene="tetL"
     CDS             2493..3869
                     /codon_start=1
                     /gene="tetL"
                     /product="tetracycline resistance protein"
                     /label=tetL
                     /protein_id="BAD00173.1"
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     terminator      3938..4024
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      3941..3984
                     /regulatory_class="terminator"
                     /gene="tetL"
                     /note="E. coli rrnB terminator T1"
     regulatory      4116..4143
                     /regulatory_class="terminator"
                     /gene="tetL"
                     /note="E. coli rrnB terminator T2"
     terminator      4116..4143
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     gene            4411..7265
                     /gene="repA"
                     /label=repA
     regulatory      4411..4439
                     /regulatory_class="promoter"
                     /gene="repA"
                     /standard_name="spac-1"
     regulatory      4411..4416
                     /regulatory_class="minus_35_signal"
                     /gene="repA"
                     /standard_name="spac-1"
     regulatory      4434..4439
                     /regulatory_class="minus_10_signal"
                     /gene="repA"
                     /standard_name="spac-1"
     protein_bind    4446..4466
                     /gene="repA"
                     /standard_name="lacO"
                     /label=lacO binding site
                     /bound_moiety="LacI"
     protein_bind    4448..4464
                     /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      6015..6044
                     /regulatory_class="promoter"
                     /gene="repA"
     regulatory      6015..6019
                     /regulatory_class="minus_35_signal"
                     /gene="repA"
     regulatory      6039..6044
                     /regulatory_class="minus_10_signal"
                     /gene="repA"
     regulatory      6062..6066
                     /regulatory_class="ribosome_binding_site"
                     /gene="repA"
     CDS             6075..7265
                     /codon_start=1
                     /gene="repA"
                     /product="RepA protein"
                     /label=repA
                     /protein_id="BAD00174.1"
                     /translation="MNNEPVKRGKKNRWELNLPIMTYVVADDWIDKLGHETFTLWLRFH
                     TWVDREDELRDYDRIPRSFENIYKKTLGISKSKFYRLIKPLWEYGLIDIIEYEESNRNS
                     TKPKNIIVYEYPLHEIERKYKPLEKLRDWDKDYNSVSKELGKTGGRPRKKDSEEEPEKK
                     PEEVTKKKRKYKLKRVIHNGFKNETVEGFKNETVEGFKNETVTVSKIKPNNYSNIFNNL
                     SNISTNVSNNLLIDDDEEIENEPTGRTINRSLLFSQEDIKQAYQFINRFSVIQLRENFS
                     FDKHFEERLVCYLWKAGISTFYTHEISKMIKKIADYEKSKKGRLNPIRDRALYMVNGLV
                     MNRASSQSEHATYKLNQYKKQKEQEKQQQEQQRSRVPFYNWLEEREEQTEGQLPTT"
     RBS             7602..7610
                     /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)"
     gene            8885..10161
                     /gene="lacI"
                     /label=lacI
     regulatory      8885..8913
                     /regulatory_class="promoter"
                     /gene="lacI"
                     /standard_name="penP promoter"
     regulatory      8885..8890
                     /regulatory_class="minus_35_signal"
                     /gene="lacI"
                     /standard_name="penP promoter"
     regulatory      8908..8913
                     /regulatory_class="minus_10_signal"
                     /gene="lacI"
                     /standard_name="penP promoter"
     regulatory      9062..9069
                     /regulatory_class="ribosome_binding_site"
                     /gene="lacI"
                     /standard_name="penP RBS"
     CDS             9079..10161
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI repressor protein"
                     /label=lacI
                     /protein_id="BAD00175.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKTAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     misc_feature    10332..10472
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(10658..11246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(11417..12277)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="BAD00176.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(11417..12277)
                     /gene="bla"
                     /label=bla
     promoter        complement(12278..12382)
                     /gene="bla"
                     /label=AmpR promoter