LOCUS       Exported                8893 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pTS1, complete sequence.
ACCESSION   KX931445
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8893)
  AUTHORS   Scott TA, Heine D, Qin Z, Wilkinson B.
  TITLE     An L-threonine transaldolase is required for 
            L-threo-beta-hydroxy-alpha-amino acid assembly during obafluorin 
            biosynthesis
  JOURNAL   Nat Commun 8, 15935 (2017)
  PUBMED    28649989
REFERENCE   2  (bases 1 to 8893)
  AUTHORS   Scott TA.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-SEP-2016) Molecular Microbiology, John Innes Centre, 
            Norwich Research Park, Norwich, Norfolk NR4 7UH, United Kingdom
REFERENCE   3  (bases 1 to 8893)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8893
                     /organism="Suicide vector pTS1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1968804"
     CDS             complement(721..1398)
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /label=tetR
                     /protein_id="AQZ26570.1"
                     /translation="MFISDKVSSMTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGV
                     QQPALYWHFRNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRD
                     GARIHAGTRPGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSD
                     AGERGGTVEQAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRK
                     GDD"
     gene            complement(721..1398)
                     /gene="tetR"
                     /label=tetR
     CDS             complement(721..1371)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             1402..2676
                     /codon_start=1
                     /gene="tetA"
                     /product="TetA"
                     /label=tetA
                     /protein_id="AQZ26571.1"
                     /translation="MSTNLSVIKNPRVQSDQRRLVRRPDVKPNIPLIVILSTVALDAVG
                     IGLIMPVLPGLLRDLVHSNDVTAHYGILLALYALVQFACAPVLGALSDRFGRRPILLVS
                     LAGATVDYAIMATAPFLWVLYIGRIVAGITGATGAVAGAYIADITDGDERARHFGFMSA
                     CFGFGMVAGPVLGGLMGGFSPHAPFFAAAALNGLNFLTGCFLLPESHKGERRPLRREAL
                     NPLASFRWARGMTVVAALMAVFFIMQLVGQVPAALWVIFGEDRFHWDATTIGISLAAFG
                     ILHSLAQAMITGPVAARLGERRALMLGMIADGTGYILLAFATRGWMAFPIMVLLASGGI
                     GMPALQAMLSRQVDEERQGQLQGSLAALTSLTSIVGPLLFTAIYAASITTWNGWAWIAG
                     AALYLLCLPALRRGLWSGAGQRADR"
     gene            1402..2676
                     /gene="tetA"
                     /label=tetA
     CDS             1477..2676
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             2679..2825
                     /codon_start=1
                     /gene="exc2"
                     /product="Exc2"
                     /label=exc2
                     /protein_id="AQZ26572.1"
                     /translation="METIGNSLRGTTQFAGTDYRSKDLTPKKSRLLADTISAVYLDGYE
                     GRQ"
     gene            2679..2825
                     /gene="exc2"
                     /label=exc2
     misc_feature    2800..3083
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     CDS             complement(3143..3376)
                     /codon_start=1
                     /gene="mbeD"
                     /product="MbeD"
                     /label=mbeD
                     /protein_id="AQZ26573.1"
                     /translation="MTELETHLLNALEQLQQDYMQRLSEWESAFVELQKMFSLTQRDNA
                     MLNERVMQLSQQVQHLSEQTERLSQLYSENWR"
     gene            complement(3143..3376)
                     /gene="mbeD"
                     /label=mbeD
     CDS             complement(3383..3901)
                     /codon_start=1
                     /gene="mbeB"
                     /product="MbeB"
                     /label=mbeB
                     /protein_id="AQZ26574.1"
                     /translation="MSNLLQTGAEFEKKLKERAESTEKMLNNEFRRLGESVSEAVTSNE
                     TKIRDAIALFTASTEESLEKHREGVKEAMMQHRRDVLKLAGNTGMMLLGIVFLLFTASG
                     GTLWYLGGRIQANLEEIRKQEETLQKLNAKTWGVEFVQDGNRKFLVLPYGKSAEVIPFQ
                     GKEWVHLKE"
     gene            complement(3383..3901)
                     /gene="mbeB"
                     /label=mbeB
     CDS             complement(4579..4917)
                     /codon_start=1
                     /gene="mbeC"
                     /product="MbeC"
                     /label=mbeC
                     /protein_id="AQZ26575.1"
                     /translation="MKRERMLTIRVTDDEHARLLERCEGKQLAVWMRRVCLGEPVARSG
                     KLPTLAPPLLRQLAAIGNNLNQTARKVNSGQWSSGDRVQVVAALMAIGDELRRLRLAVR
                     EQGARDDS"
     gene            complement(4579..4917)
                     /gene="mbeC"
                     /label=mbeC
     CDS             4870..5157
                     /codon_start=1
                     /gene="rop"
                     /product="Rop"
                     /label=rop
                     /protein_id="AQZ26576.1"
                     /translation="MFIISNPYCEHPLAFHRYHYPMNRNPPYTEASVTKQEKTALNMAR
                     FIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL"
     gene            4870..5157
                     /gene="rop"
                     /label=rop
     oriT            5242..5330
                     /note="oriT"
     rep_origin      5758..5993
                     /label=oriV
                     /note="oriV"
     CDS             6369..6710
                     /codon_start=1
                     /gene="colE1"
                     /product="colicin E1 immunity protein"
                     /label=colE1
                     /protein_id="AQZ26577.1"
                     /translation="MSLRYYIKNILFGLYCTLIYIYLITKNSEGYYFLVSDKMLYAIVI
                     STILCPYSKYAIEYIAFNFIKKDFFERRKNLNNAPVAKLNLFMLYNLLCLVLAIPFGLL
                     GLFISIKNN"
     gene            6369..6710
                     /gene="colE1"
                     /label=colE1
     CDS             complement(6887..8308)
                     /codon_start=1
                     /gene="sacB"
                     /product="SacB"
                     /label=sacB
                     /protein_id="AQZ26578.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(6887..8308)
                     /gene="sacB"
                     /label=sacB
     promoter        complement(8309..8754)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     misc_feature    8777..8833
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    8783..8888
                     /label=multiple cloning site
                     /note="multiple cloning site"