LOCUS       40924_32658        4941 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMTL83353, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4941)
  AUTHORS   Heap JT, Pennington OJ, Cartman ST, Minton NP.
  TITLE     A modular system for Clostridium shuttle plasmids
  JOURNAL   J. Microbiol. Methods 78 (1), 79-85 (2009)
  PUBMED    19445976
REFERENCE   2  (bases 1 to 4941)
  AUTHORS   Pennington OJ, Heap JT, Cartman ST, Minton NP.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAR-2009) Centre for Biomolecular Sciences, The 
            University of Nottingham, University Park, Nottingham NG7 2RD, 
            United Kingdom
REFERENCE   3  (bases 1 to 4941)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4941)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Microbiol. Methods"; date: "2009"; volume: "78"; issue: "1"; pages: 
            "79-85"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-MAR-2009) Centre for Biomolecular Sciences, The University of 
            Nottingham, University Park, Nottingham NG7 2RD, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      9..62
                     /note="terminator derived from CD0164 gene of Clostridium 
                     difficile"
                     /regulatory_class="terminator"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      89..288
                     /note="promoter derived from fdx gene of Clostridium
                     sporogenes"
                     /regulatory_class="promoter"
     RBS             274..282
                     /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)"
     primer_bind     complement(393..409)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      557..598
                     /note="terminator derived from fdx gene of Clostridium 
                     pasteurianum"
                     /regulatory_class="terminator"
     CDS             1325..1642
                     /codon_start=1
                     /gene="repH"
                     /product="putative plasmid replication protein"
                     /label=repH
                     /protein_id="ACR43901.1"
                     /translation="MKRRAYMKTCKNCKEFIKDTEICKIHSLMIHDKTVATYCSKYNES
                     RCLHKGKVQCINCSKMNRYGWCAIKMRCFTEEEQKKERTCIKYYARSFKKAHVKKSKKK
                     K"
     gene            1325..1642
                     /gene="repH"
                     /label=repH
     CDS             2434..3186
                     /codon_start=1
                     /gene="aad9"
                     /product="spectinomycin resistance protein"
                     /label=aad9
                     /protein_id="ACR43899.1"
                     /translation="MNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNSDLDFL
                     VVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYG
                     EWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMD
                     SSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERILLAVR
                     SYLGENIEWTNENVNLTINYLNNRLKKL"
     gene            2434..3186
                     /gene="aad9"
                     /label=aad9
     rep_origin      3385..3973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     oriT            4294..4403
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             4436..4804
                     /label=traJ
                     /note="oriT-recognizing protein"
ORIGIN
        1 cctgcaggat aaaaaaattg tagataaatt ttataaaata gttttatcta caattttttt
       61 atcaggaaac agctatgacc gcggccgcgt gtagtagcct gtgaaataag taaggaaaaa
      121 aaagaagtaa gtgttatata tgatgattat tttgtagatg tagataggat aatagaatcc
      181 atagaaaata taggttatac agttatataa aaattacttt aaaaattaat aaaaacatgg
      241 taaaatataa atcgtataaa gttgtgtaat ttttaaggag gtgtgttaca tatgaccatg
      301 attacgaatt cgagctcggt acccggggat cctctagagt cgacgtcacg cgtccatgga
      361 gatctcgagg cctgcagaca tgcaagcttg gcactggccg tcgttttaca acgtcgtgac
      421 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
      481 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
      541 ggcgaatggc gctagcataa aaataagaag cctgcatttg caggcttctt atttttatgg
      601 cgcgccgcca ttattttttt gaacaattga caattcattt cttatttttt attaagtgat
      661 agtcaaaagg cataacagtg ctgaatagaa agaaatttac agaaaagaaa attatagaat
      721 ttagtatgat taattatact catttatgaa tgtttaattg aatacaaaaa aaaatacttg
      781 ttatgtattc aattacgggt taaaatatag acaagttgaa aaatttaata aaaaaataag
      841 tcctcagctc ttatatatta agctaccaac ttagtatata agccaaaact taaatgtgct
      901 accaacacat caagccgtta gagaactcta tctatagcaa tatttcaaat gtaccgacat
      961 acaagagaaa cattaactat atatattcaa tttatgagat tatcttaaca gatataaatg
     1021 taaattgcaa taagtaagat ttagaagttt atagcctttg tgtattggaa gcagtacgca
     1081 aaggcttttt tatttgataa aaattagaag tatatttatt ttttcataat taatttatga
     1141 aaatgaaagg gggtgagcaa agtgacagag gaaagcagta tcttatcaaa taacaaggta
     1201 ttagcaatat cattattgac tttagcagta aacattatga cttttatagt gcttgtagct
     1261 aagtagtacg aaagggggag ctttaaaaag ctccttggaa tacatagaat tcataaatta
     1321 atttatgaaa agaagggcgt atatgaaaac ttgtaaaaat tgcaaagagt ttattaaaga
     1381 tactgaaata tgcaaaatac attcgttgat gattcatgat aaaacagtag caacctattg
     1441 cagtaaatac aatgagtcaa gatgtttaca taaagggaaa gtccaatgta ttaattgttc
     1501 aaagatgaac cgatatggat ggtgtgccat aaaaatgaga tgttttacag aggaagaaca
     1561 gaaaaaagaa cgtacatgca ttaaatatta tgcaaggagc tttaaaaaag ctcatgtaaa
     1621 gaagagtaaa aagaaaaaat aatttattta ttaatttaat attgagagtg ccgacacagt
     1681 atgcactaaa aaatatatct gtggtgtagt gagccgatac aaaaggatag tcactcgcat
     1741 tttcataata catcttatgt tatgattatg tgtcggtggg acttcacgac gaaaacccac
     1801 aataaaaaaa gagttcgggg tagggttaag catagttgag gcaactaaac aatcaagcta
     1861 ggatatgcag tagcagaccg taaggtcgtt gtttaggtgt gttgtaatac atacgctatt
     1921 aagatgtaaa aatacggata ccaatgaagg gaaaagtata atttttggat gtagtttgtt
     1981 tgttcatcta tgggcaaact acgtccaaag ccgtttccaa atctgctaaa aagtatatcc
     2041 tttctaaaat caaagtcaag tatgaaatca taaataaagt ttaattttga agttattatg
     2101 atattatgtt tttctattaa aataaattaa gtatatagaa tagtttaata atagtatata
     2161 cttaatgtga taagtgtctg acagtgtcac agaaaggatg attgttatgg attataagcg
     2221 gccggcccaa tgaataggtt tacacttact ttagttttat ggaaatgaaa gatcatatca
     2281 tatataatct agaataaaat taactaaaat aattattatc tagataaaaa atttagaagc
     2341 caatgaaatc tataaataaa ctaaattaag tttatttaat taacaactat ggatataaaa
     2401 taggtactaa tcaaaatagt gaggaggata tatttgaata catacgaaca aattaataaa
     2461 gtgaaaaaaa tacttcggaa acatttaaaa aataacctta ttggtactta catgtttgga
     2521 tcaggagttg agagtggact aaaaccaaat agtgatcttg actttttagt cgtcgtatct
     2581 gaaccattga cagatcaaag taaagaaata cttatacaaa aaattagacc tatttcaaag
     2641 aaaataggag ataaaagcaa cttacgatat attgaattaa caattattat tcagcaagaa
     2701 atggtaccgt ggaatcatcc tcccaaacaa gaatttattt atggagaatg gttacaagag
     2761 ctttatgaac aaggatacat tcctcagaag gaattaaatt cagatttaac cataatgctt
     2821 taccaagcaa aacgaaaaaa taaaagaata tacggaaatt atgacttaga ggaattacta
     2881 cctgatattc cattttctga tgtgagaaga gccattatgg attcgtcaga ggaattaata
     2941 gataattatc aggatgatga aaccaactct atattaactt tatgccgtat gattttaact
     3001 atggacacgg gtaaaatcat accaaaagat attgcgggaa atgcagtggc tgaatcttct
     3061 ccattagaac atagggagag aattttgtta gcagttcgta gttatcttgg agagaatatt
     3121 gaatggacta atgaaaatgt aaatttaact ataaactatt taaataacag attaaaaaaa
     3181 ttataaaaaa attgaaaaaa tggtggaaac acttttttca atttttttgt tttattattt
     3241 aatatttggg aaatattcat tctaattggt aatcagattt tagaagttta aactcctttt
     3301 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3361 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3421 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3481 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3541 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3601 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3661 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3721 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3781 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3841 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3901 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3961 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4021 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4081 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4141 cgaggaagcg gaagagcgcc caatacgcag ggccccctgc ttcggggtca ttatagcgat
     4201 tttttcggta tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt
     4261 agactttcct tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc
     4321 cgcgagcggg tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa
     4381 tcctgctctg cgaggctggc cggctaccgc cggcgtaaca gatgagggca agcggatggc
     4441 tgatgaaacc aagccaacca ggaagggcag cccacctatc aaggtgtact gccttccaga
     4501 cgaacgaaga gcgattgagg aaaaggcggc ggcggccggc atgagcctgt cggcctacct
     4561 gctggccgtc ggccagggct acaaaatcac gggcgtcgtg gactatgagc acgtccgcga
     4621 gctggcccgc atcaatggcg acctgggccg cctgggcggc ctgctgaaac tctggctcac
     4681 cgacgacccg cgcacggcgc ggttcggtga tgccacgatc ctcgccctgc tggcgaagat
     4741 cgaagagaag caggacgagc ttggcaaggt catgatgggc gtggtccgcc cgagggcaga
     4801 gccatgactt ttttagccgc taaaacggcc ggggggtgcg cgtgattgcc aagcacgtcc
     4861 ccatgcgctc catcaagaag agcgacttcg cggagctggt gaagtacatc accgacgagc
     4921 aaggcaagac cgatcgggcc c
//
