LOCUS       40924_44314        8893 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pTS1, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8893)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            8, 15935 (2017)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-SEP-2016) Molecular Microbiology, John Innes Centre, Norwich 
            Research Park, Norwich, Norfolk NR4 7UH, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(724..1371)
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
     CDS             1477..2673
                     /label=TcR
                     /note="tetracycline efflux protein"
     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    complement(2800..3083)
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     CDS             complement(3039..4589)
                     /gene="mbeA"
                     /label=mbeA
                     /note="DNA relaxase MbeA from Escherichia coli. Accession#:
                     P13658"
     CDS             complement(4582..4926)
                     /gene="mbeC"
                     /label=mbeC
                     /note="Mobilization protein MbeC from Escherichia coli. 
                     Accession#: P13657"
     CDS             4966..5154
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     oriT            5242..5330
                     /note="oriT"
     rep_origin      complement(5574..6164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     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(6890..8308)
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     promoter        complement(8309..8754)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     misc_feature    8777..8833
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
ORIGIN
        1 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
       61 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
      121 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
      181 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
      241 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
      301 acgctgactt gacgggacgg cggctttgtt gaataaatcg aacttttgct gagttgaagg
      361 atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa
      421 tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc actttctggc
      481 tggatgatgg ggcgattcag gcctggtatg agtcagcaac accttcttca cgaggcagac
      541 ctcagcgcca gaaggccgcc agagaggccg agcgcggccg tgaggcttgg acgctagggc
      601 agggcatgaa aaagcccgta gcgggctgct acgggcgtct gacgcggtgg aaagggggag
      661 gggatgttgt ctacatggct ctgctgtagt gagtgggttg cgctccggca gcggtcctga
      721 tcaatcgtca ccctttctcg gtccttcaac gttcctgaca acgagcctcc ttttcgccaa
      781 tccatcgaca atcaccgcga gtccctgctc gaacgctgcg tccggaccgg cttcgtcgaa
      841 ggcgtctatc gcggcccgca acagcggcga gagcggagcc tgttcaacgg tgccgccgcg
      901 ctcgccggca tcgctgtcgc cggcctgctc ctcaagcacg gccccaacag tgaagtagct
      961 gattgtcatc agcgcattga cggcgtcccc ggccgaaaaa cccgcctcgc agaggaagcg
     1021 aagctgcgcg tcggccgttt ccatctgcgg tgcgcccggt cgcgtgccgg catggatgcg
     1081 cgcgccatcg cggtaggcga gcagcgcctg cctgaagctg cgggcattcc cgatcagaaa
     1141 tgagcgccag tcgtcgtcgg ctctcggcac cgaatgcgta tgattctccg ccagcatggc
     1201 ttcggccagt gcgtcgagca gcgcccgctt gttcctgaag tgccagtaaa gcgccggctg
     1261 ctgaaccccc aaccgttccg ccagtttgcg tgtcgtcaga ccgtctacgc cgacctcgtt
     1321 caacaggtcc agggcggcac ggatcactgt attcggctgc aactttgtca tgcttgacac
     1381 tttatcactg ataaacataa tatgtccacc aacttatcag tgataaagaa tccgcgcgtt
     1441 caatcggacc agcggaggct ggtccggagg ccagacgtga aacccaacat acccctgatc
     1501 gtaattctga gcactgtcgc gctcgacgct gtcggcatcg gcctgattat gccggtgctg
     1561 ccgggcctcc tgcgcgatct ggttcactcg aacgacgtca ccgcccacta tggcattctg
     1621 ctggcgctgt atgcgttggt gcaatttgcc tgcgcacctg tgctgggcgc gctgtcggat
     1681 cgtttcgggc ggcggccaat cttgctcgtc tcgctggccg gcgccactgt cgactacgcc
     1741 atcatggcga cagcgccttt cctttgggtt ctctatatcg ggcggatcgt ggccggcatc
     1801 accggggcga ctggggcggt agccggcgct tatattgccg atatcactga tggcgatgag
     1861 cgcgcgcggc acttcggctt catgagcgcc tgtttcgggt tcgggatggt cgcgggacct
     1921 gtgctcggtg ggctgatggg cggtttctcc ccccacgctc cgttcttcgc cgcggcagcc
     1981 ttgaacggcc tcaatttcct gacgggctgt ttccttttgc cggagtcgca caaaggcgaa
     2041 cgccggccgt tacgccggga ggctctcaac ccgctcgctt cgttccggtg ggcccggggc
     2101 atgaccgtcg tcgccgccct gatggcggtc ttcttcatca tgcaacttgt cggacaggtg
     2161 ccggccgcgc tttgggtcat tttcggcgag gatcgctttc actgggacgc gaccacgatc
     2221 ggcatttcgc ttgccgcatt tggcattctg cattcactcg cccaggcaat gatcaccggc
     2281 cctgtagccg cccggctcgg cgaaaggcgg gcactcatgc tcggaatgat tgccgacggc
     2341 acaggctaca tcctgcttgc cttcgcgaca cggggatgga tggcgttccc gatcatggtc
     2401 ctgcttgctt cgggtggcat cggaatgccg gcgctgcaag caatgttgtc caggcaggtg
     2461 gatgaggaac gtcaggggca gctgcaaggc tcactggcgg cgctcaccag cctgacctcg
     2521 atcgtcggac ccctcctctt cacggcgatc tatgcggctt ctataacaac gtggaacggg
     2581 tgggcatgga ttgcaggcgc tgccctctac ttgctctgcc tgccggcgct gcgtcgcggg
     2641 ctttggagcg gcgcagggca acgagccgat cgctgatcgt ggaaacgata ggaaactcac
     2701 tccggggcac aactcaattt gcgggtactg attaccgcag caaagacctt accccgaaaa
     2761 aatccaggct gctggctgac acgatttctg cggtttatct cgatggctac gagggcagac
     2821 agtaagtgga tttaccataa tcccttaatt gtacgcaccg ctaaaacgcg ttcagcgcga
     2881 tcacggcagc agacaggtaa aaatggcaac aaaccacccg aaaaactgcc gcgatcgcgc
     2941 ctgataaatt ttaaccgtat gaatacctat gcaaccagag ggtacaggcc acattacccc
     3001 cacttaatcc actgaagctg ccatttttca tggtttcacc atcccagcga agggccatcc
     3061 agcgtgcgtt cctgtatttc cggctgacgc tcccgttcca gggataacac atgttcgcgc
     3121 tcctgtatca gccgttcctc tcttatctcc agttctcgct gtataactgg ctcaagcgtt
     3181 ctgtctgctc gctcaagtgc tgcacctgct gactcaactg catgacccgc tcgttcagca
     3241 tcgcgttgtc ccgttgcgta agcgaaaaca tcttctgcaa ttccacgaag gcgctctccc
     3301 attcgctcag ccgctgcata tagtcctgtt gcagctgctc taaggcgttc agcaaatgtg
     3361 tttccagctc tgtcactctg tgtcactcct tcagatgtac ccactctttc ccctgaaagg
     3421 gaatcacctc cgctgatttc ccgtacggaa ggacaaggaa tttcctgttc ccgtcctgca
     3481 caaactccac gccccatgtc ttcgcgttca gtttctgcaa tgtctcttcc tgcttcctga
     3541 tttcttccag gttcgcctgt atcctccctc caagatacca gagcgtcccg ccactcgcgg
     3601 taaacaggag aaagactatc cccagtaaca tcatgcccgt attccctgcc agctttaaca
     3661 cgtccctcct gtgctgcatc atcgcctctt tcaccccttc ccggtgtttt tccagcgatt
     3721 cctctgtcga ggctgtgaac agggctatag cgtctctgat tttcgtctcg tttgatgtca
     3781 cagcctcgct tacagattcg ccgagcctcc tgaactcgtt gttcagcatt ttctctgtag
     3841 attcggctct ctctttcagc tttttctcga actccgcgcc cgtctgcaaa agattgctca
     3901 taaaatgctc ctttcagcct gatattcttc ccgccgttcg gatctgcaat gctgatactg
     3961 cttcgcgtca ccctgaccac ttccagcccc gcctcagtga gcgcctgaat cacatcctga
     4021 cggcctttta tctctccggc atggtaaagt gcatctatac ctcgcgtgac gccctcagca
     4081 agcgcctgtt tcgtttcagg caggttatca gggagtgtca gcgtcctgcg gttctccggg
     4141 gcgttcgggt catgcagccc gtaatggtga tttaccagcg tctgccaggc atcaattcta
     4201 ggcctgtctg cgcggtcgta gtacggctgg aggcgttttc cggtctgtag ctccatgttc
     4261 ggaatgacaa aattcagctc aagccgtccc ttgtcctggt gctccaccca caggatgctg
     4321 tactgatttt tttcgagacc gggcatcagt acacgctcaa agctcgccat cactttttca
     4381 cgtcctcccg gcggcagctc cttctccgcg aacgacagaa caccggacgt gtatttcttc
     4441 gcaaatggcg tggcatcgat gagttcccgg acttcttccg gattaccctg aagcaccgtt
     4501 gcgccttcgc ggttacgctc cctccccagc aggtaatcaa ccggaccact gccaccacct
     4561 tttcccctgg catgaaattt aactatcatc ccgcgccccc tgttccctga cagccagacg
     4621 cagccggcgc agctcatccc cgatggccat cagtgcggcc accacctgaa cccggtcacc
     4681 ggaagaccac tgcccgctgt tcaccttacg ggctgtctga ttcaggttat ttccgatggc
     4741 ggccagctga cgcagtaacg gcggtgccag tgtcggcagt tttccggaac gggcaaccgg
     4801 ctcccccagg cagacccgcc gcatccatac cgccagttgt ttaccctcac agcgttcaag
     4861 taaccgggca tgttcatcat cagtaacccg tattgtgagc atcctctcgc gtttcatcgg
     4921 tatcattacc ccatgaacag aaatccccct tacacggagg catcagtgac taaacaggaa
     4981 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct gctggagaag
     5041 ctcaacgaac tggacgcaga tgaacaggcc gatatttgtg aatcgcttca cgaccacgcc
     5101 gatgagcttt accgcagctg cctcgcacgt ttcggggatg acggtgaaaa cctctgacac
     5161 atgcagctcc cggagacggt cacagcttgt ctgtgagcgg atgccgggag ctgacaagcc
     5221 cgtcagggcg cgtcagcagg ttttagcggg tgtcggggcg cagccctgac ccagtcacgt
     5281 agcgatagcg gagtgtatac tggcttaacc atgcggcatc agtgcggatt gtatgaaaag
     5341 tacgccatgc cgggtgtgaa atgccgcaca gatgcgtaag gagaaaatgc acgtccaggc
     5401 gcttttccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg
     5461 tactgactca cacaaaaacg gtaacacagt tatccacaga atcaggggat aaggccggaa
     5521 agaacatgtg agcaaaagac caggaacagg aagaaggcca cgtagcaggc gtttttccat
     5581 aggctccgcc cccctgacga gcatcacaaa aatagacgct caagtcagag gtggcgaaac
     5641 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5701 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5761 ctttctcata gctcacgctg ttggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5821 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5881 cttgagtcca acccggtaag gcacgcctta acgccactgg cagcagccac tggtaaccgg
     5941 attagcagag cgatgatggc acaaacggtg ctacagagtt cttgaagtag tggcccgact
     6001 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6061 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgttggtagc ggtggttttt
     6121 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttaatct
     6181 tttctactga accgcgatcc ccgtcagttt agaagaggag gatggtgcga tggtccctcc
     6241 ctgaacatca cgtatatagt tagcctgaca tccaacaagg aggtttatcg cgaatattcc
     6301 cacaaaaaat cttttcctca taactcgatc cttataaaat gaaaagaata tatggcgagg
     6361 tttaatttat gagcttaaga tactacataa aaaatatttt atttggcctg tactgcacac
     6421 ttatatatat ataccttata acaaaaaaca gcgaagggta ttatttcctt gtgtcagata
     6481 agatgctata tgcaatagtg ataagcacta ttctatgtcc atattcaaaa tatgctattg
     6541 aatacatagc ttttaacttc ataaagaaag attttttcga aagaagaaaa aacctaaata
     6601 acgcccccgt agcaaaatta aacctattta tgctatataa tctactttgt ttggtcctag
     6661 caatcccatt tggattgcta ggacttttta tatcaataaa gaataattaa atccctaaca
     6721 cctcatttat agtattaagt ttattcttat caatatagga gcatagaatt cggaatacgg
     6781 ttagccattt gcctgctttt atatagttca tatatgggat tcacctttat gttgataaga
     6841 aataaaagaa aatgccaata ggatatcggc attttctttt gcgtttttat ttgttaactg
     6901 ttaattgtcc ttgttcaagg atgctgtctt tgacaacaga tgttttcttg cctttgatgt
     6961 tcagcaggaa gctaggcgca aacgttgatt gtttgtctgc gtagaatcct ctgtttgtca
     7021 tatagcttgt aatcacgaca ttgtttcctt tcgcttgagg tacagcgaag tgtgagtaag
     7081 taaaggttac atcgttagga tcaagatcca tttttaacac aaggccagtt ttgttcagcg
     7141 gcttgtatgg gccagttaaa gaattagaaa cataaccaag catgtaaata tcgttagacg
     7201 taatgccgtc aatcgtcatt tttgatccgc gggagtcagt gaacagatac catttgccgt
     7261 tcattttaaa gacgttcgcg cgttcaattt catctgttac tgtgttagat gcaatcagcg
     7321 gtttcatcac ttttttcagt gtgtaatcat cgtttagctc aatcataccg agagcgccgt
     7381 ttgctaactc agccgtgcgt tttttatcgc tttgcagaag tttttgactt tcttgacgga
     7441 agaatgatgt gcttttgcca tagtatgctt tgttaaataa agattcttcg ccttggtagc
     7501 catcttcagt tccagtgttt gcttcaaata ctaagtattt gtggccttta tcttctacgt
     7561 agtgaggatc tctcagcgta tggttgtcgc ctgagctgta gttgccttca tcgatgaact
     7621 gctgtacatt ttgatacgtt tttccgtcac cgtcaaagat tgatttataa tcctctacac
     7681 cgttgatgtt caaagagctg tctgatgctg atacgttaac ttgtgcagtt gtcagtgttt
     7741 gtttgccgta atgtttaccg gagaaatcag tgtagaataa acggattttt ccgtcagatg
     7801 taaatgtggc tgaacctgac cattcttgtg tttggtcttt taggatagaa tcatttgcat
     7861 cgaatttgtc gctgtcttta aagacgcggc cagcgttttt ccagctgtca atagaagttt
     7921 cgccgacttt ttgatagaac atgtaaatcg atgtgtcatc cgcattttta ggatctccgg
     7981 ctaatgcaaa gacgatgtgg tagccgtgat agtttgcgac agtgccgtca gcgttttgta
     8041 atggccagct gtcccaaacg tccaggcctt ttgcagaaga gatattttta attgtggacg
     8101 aatcgaactc aggaacttga tatttttcat ttttttgctg ttcagggatt tgcagcatat
     8161 catggcgtgt aatatgggaa atgccgtatg tttccttata tggcttttgg ttcgtttctt
     8221 tcgcaaacgc ttgagttgcg cctcctgcca gcagtgcggt agtaaaggtt aatactgttg
     8281 cttgttttgc aaactttttg atgttcatcg ttcatgtctc cttttttatg tactgtgtta
     8341 gcggtctgct tcttccagcc ctcctgtttg aagatggcaa gttagttacg cacaataaaa
     8401 aaagacctaa aatatgtaag gggtgacgcc aaagtataca ctttgccctt tacacatttt
     8461 aggtcttgcc tgctttatca gtaacaaacc cgcgcgattt acttttcgac ctcattctat
     8521 tagactctcg tttggattgc aactggtcta ttttcctctt ttgtttgata gaaaatcata
     8581 aaaggatttg cagactacgg gcctaaagaa ctaaaaaatc tatctgtttc ttttcattct
     8641 ctgtattttt tatagtttct gttgcatggg cataaagttg cctttttaat cacaattcag
     8701 aaaatatcat aatatctcat ttcactaaat aatagtgaac ggcaggtata tgtgatgggt
     8761 taaaaaggat cgatccgaat tcgagctcgg tacccgggga tcctctagag tcgacctgca
     8821 ggcatgcaag cttcctaggt agtcaattgg tcaccatggt tggctagcac ctctcgaggc
     8881 atcatatgag ctt
//
