LOCUS       40924_32668        7371 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMTL84422, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7371)
  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 7371)
  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 7371)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7371)
  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..7371
                     /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..238
                     /note="promoter derived from thl gene of Clostridium 
                     acetobutylicum"
                     /regulatory_class="promoter"
     primer_bind     complement(343..359)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      507..548
                     /note="terminator derived from fdx gene of Clostridium 
                     pasteurianum"
                     /regulatory_class="terminator"
     CDS             713..1360
                     /codon_start=1
                     /product="putative plasmid replication protein"
                     /label=putative plasmid replication protein
                     /note="orfB"
                     /protein_id="ACR43910.1"
                     /translation="MNKKLEKPFVYKREYDLTGYDVEILQKYELEQAIYVYVGSSCAYN
                     MRARSSKWRYHIRTNNKSICCNIKNFIHNLELFYKMELKLSDNIINDKLYYSNIAEFEE
                     FETLEKAREVESTIISQYQFLDSINHMLKQKIILLSNKDSVLNITKNGNTNYLKVKNKY
                     IEKHKNKPIMRYHINCQFNTDGSVKSITQEFEPILELNKKNTLSRPSRVFLK"
     CDS             1635..3272
                     /codon_start=1
                     /gene="repA"
                     /product="plasmid replication protein"
                     /label=repA
                     /protein_id="ACR43908.1"
                     /translation="MEQLDSKYKLKKFLMAVFRDGIGQGNNLIDNEYVRVFQNNKSNSK
                     QLELGEEFKEYSKTTFFKNIDDIVEFTFAKNIYYENTFFNLCTTDGKAGTNENLINRYA
                     LGFDFDKKELGQGFNYKDIINLFTKIGLHYHILVDSGNGFHVYVLINKTNNIKLVSEVT
                     NTLINKLGADKQANLSTQVLRVPYTYNIKNTTKQVKIIHQDKNIYRYDIEKLAKKYCKD
                     VKTVGNTNTKYILDSKLPNCIVDILKNGSKDGHKNLDLQKIVVTLRLRNKSLSQVISVA
                     REWNYISQNSLSNSELEYQVKYMYEKLKTVNFGCTGCEFNSDCWNKIESDFIYSDEDTL
                     FNMPHKHSKDLKYKNRKGVKIMTGNQLFIYNVLLNNKDRELNIDDIMELITYKRKKKVK
                     NIVMSEKTLRETLKELQHNDYITKTKGVTKLGIKDTYNVKEVRCNIDKQYTISYFVTMA
                     VIWGIISTEELRLYTHMRYKQDLLVKDDKIKGNILRINQEELAKDLGVTQQRISNMIES
                     LLDTKILDVWETKINDRGFMYYTYRLNK"
     gene            1635..3272
                     /gene="repA"
                     /label=repA
     rep_origin      complement(3478..3717)
                     /direction=LEFT
                     /label=oriV
                     /note="oriV"
     CDS             4620..5882
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline resistance protein"
                     /label=tetA
                     /protein_id="ACR43909.1"
                     /translation="MVNKLSAYKTYLLFSAITAMCFSLVATVMMVYHIEIVHLNPLQLI
                     LVGTTLELACFIFEIPTAIVADVYSRKLSIVIGGVLTGVGFILEGSISSFVFVLVAQIV
                     WGLGSTFISGSLEAWIAEEEKNKDLDEIYIKGAQAGQIGAFIGIVLSTVIANFSVRLPI
                     IVSGVLFIILALFLWLYMPENNFKPSAPGDLNTFKKMVYTFKSGLKFVKSKSIIMILLA
                     VTLFYGLSSEGYDRLSNAHFLQDTTLPKLGNLSSVTWFGIFGILGMILSFIVMHFMAKN
                     LKNEDNRKNGKLLLCINILYISSMLIFALTRNFSLMLIAYLATNTFRIINKPIFSAWLN
                     GHIDDNSRATVLSINGQMNSLGQILGGPIIGIIATNISVSIGIVCTSLLVTPVLVLYIV
                     AMIIDKKVDDRVGGIDYEENN"
     gene            4620..5882
                     /gene="tetA"
                     /label=tetA
     rep_origin      5931..6476
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     oriT            6724..6833
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             6866..7234
                     /label=traJ
                     /note="oriT-recognizing protein"
ORIGIN
        1 cctgcaggat aaaaaaattg tagataaatt ttataaaata gttttatcta caattttttt
       61 atcaggaaac agctatgacc gcggccgctt tttaacaaaa tatattgata aaaataataa
      121 tagtgggtat aattaagttg ttagagaaaa cgtataaatt agggataaac tatggaactt
      181 atgaaataga ttgaaatggt ttatctgtta ccccgtatca aaatttagga ggttagttca
      241 tatgaccatg attacgaatt cgagctcggt acccggggat cctctagagt cgacgtcacg
      301 cgtccatgga gatctcgagg cctgcagaca tgcaagcttg gcactggccg tcgttttaca
      361 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
      421 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
      481 cagcctgaat ggcgaatggc gctagcataa aaataagaag cctgcatttg caggcttctt
      541 atttttatgg cgcgcccgcc cttaagtcta aaaattaggg gagatgtaag gatttgggaa
      601 aaatagaaga tgttataatc ataaatatgg tattcgtagg cttaaagtca aaaaggaggt
      661 gaaatataaa tagattttta gctaaattaa gtaagaaata ggaggagatt tattgaacaa
      721 aaaattagaa aaaccatttg tatataagag agagtacgat ttgactggat atgatgttga
      781 aattttacaa aaatatgagt tagaacaagc aatatatgtt tatgttggga gtagttgtgc
      841 atataacatg agagctagaa gtagtaaatg gagataccat ataagaacaa ataataagtc
      901 tatatgttgt aacattaaaa attttataca taacttggaa ttgttttata aaatggaatt
      961 aaagttgtca gataatatta ttaatgataa gctatactat agcaatatag cagagtttga
     1021 agaatttgaa acactagaaa aagctagaga ggtagaaagt actattataa gtcaatatca
     1081 atttttagat tctataaatc acatgttaaa acaaaaaata attttattga gtaataagga
     1141 tagtgtgtta aacataacta aaaatggaaa tacaaattat ttgaaagtaa aaaataaata
     1201 catagaaaaa cataagaaca agccaataat gagataccat atcaactgtc aattcaatac
     1261 agatggaagt gtcaaaagta ttacacagga gtttgaacca atattggaat taaacaaaaa
     1321 aaatacccta agccgaccaa gcagagtatt tttaaaataa tattttaaga taacaacaaa
     1381 atgagataat actactagac aatgacaact caactaccaa ttgagtttat ggagctacca
     1441 actccaatat cggtctaact gattaagtat ctgtagttat ataataatat tgctatcaat
     1501 tttagcatct taacaatatt attatacata ctaagctaaa attattcaat agttgtaaaa
     1561 gttgattagt caataagtat atatttaatg tagtgttatc tcttaaaaaa actagataag
     1621 gagataataa atatatggaa caattagatt caaaatataa gttgaaaaaa tttctaatgg
     1681 cagtatttag agatggtata ggacaaggaa ataatcttat tgataatgaa tatgttagag
     1741 tatttcaaaa taataaaagt aatagtaaac aattagaact cggagaagaa tttaaagaat
     1801 atagtaaaac aacttttttt aaaaatatag atgatatagt agaatttacc ttcgcaaaaa
     1861 atatttatta tgaaaataca ttttttaacc tatgtactac tgatggaaaa gcaggaacca
     1921 atgaaaactt aataaataga tatgcattag gatttgattt tgacaaaaaa gaattaggac
     1981 aaggttttaa ttataaagat ataattaatt tatttactaa gataggatta cattatcata
     2041 tcctagttga tagtggaaat ggattccatg tttatgtgct aattaataaa actaataaca
     2101 ttaagttagt atcagaagtt acaaatacat taataaataa attgggtgca gataaacaag
     2161 caaatttatc tactcaagta ttaagagtac cttatacata taatattaaa aatactacta
     2221 aacaagtaaa aataatacac caagacaaaa atatatatag atatgacata gaaaagttag
     2281 ctaaaaaata ttgcaaagat gtaaaaacag taggtaatac taatacaaaa tatatattag
     2341 atagtaagct accaaattgt atagtagata ttttaaaaaa tggtagtaaa gatggacata
     2401 aaaacctaga tttgcaaaaa atagttgtga ctttaagatt gaggaataaa agtttaagtc
     2461 aagtaatatc cgttgctaga gaatggaact atatatcaca aaatagtctt tcaaatagtg
     2521 agctagaata tcaagtcaag tatatgtatg agaaacttaa aacggttaat tttggttgta
     2581 ctggttgtga gtttaatagt gattgttgga ataaaataga atcagatttt atatatagtg
     2641 atgaagatac tttgttcaat atgccacata agcactcaaa ggatttgaaa tataagaata
     2701 ggaaaggggt taaaataatg actggtaatc aattgtttat ctataatgtg ttacttaaca
     2761 ataaagatag agaattaaac atagacgata taatggagct gataacctat aaacgtaaga
     2821 agaaagttaa aaacattgtt atgagtgaaa agacattaag agaaacatta aaagaacttc
     2881 aacataatga ttatattaca aaaacaaaag gtgttacaaa gctaggaata aaagatacat
     2941 acaatgtaaa agaagttaga tgtaatatag ataaacaata tactattagt tactttgtta
     3001 ccatggcagt aatttgggga ataatttcaa ctgaagaatt aagattatat actcacatga
     3061 gatataagca agatttattg gtcaaagatg ataaaataaa aggaaatata ttaagaatta
     3121 atcaagagga attagcaaaa gatttaggag taacacagca aagaatttca aatatgatag
     3181 aatctttatt agatactaaa attttagatg tatgggaaac taaaataaat gatagaggat
     3241 ttatgtacta tacatataga ttaaacaagt agatttttga taggattaga attgattttc
     3301 tagtcctatt tttatgcaaa aaaactaatt aataaaaatt tcttttggta aaataattgt
     3361 acgagaattg caaaaaaaaa atggcatcaa agtattgaaa ttaagccgtt ttaaaaattt
     3421 cttttggtaa aataattcta catatatatg tagtatatat atatatgttt tttagagaat
     3481 gtataactag aatatagagc tagaatatag agaatgtata actagaatat agagctagaa
     3541 tatagagaat gtataactag aatatagagc tagaatatag agaatgtata actagaatat
     3601 agagctagaa tatagagaat gtataactag aatatagagc tagaatatag agaatgtata
     3661 actagaatat agagctagaa tatagagaat gtataactag aatatagagc tagaatccta
     3721 attagtaggt gcttttttaa aacaagttaa aaatcaaaaa tagtattagt aagcattgga
     3781 aatgctagat tctaaaatag aaaagtaaaa aattggtgca ctatctaaac ttatctatat
     3841 cgctttttcc gtcgtttggt tctctagtta cgatacaggg gatatgctta tattgagtta
     3901 tagtactaat cagtgcttaa tatagttaat aaaattatag ttaccatagt ttagtaacta
     3961 tgatgtatgt tagttagaaa cttgcatttc ggccggccac tctataattt agagtctttt
     4021 ttgattttaa ttaaatttgg ataaatataa atataaaaat attgaaaaaa aataaaaaaa
     4081 aatatataat gtagttatgc taaaatataa acatgaaaaa tataaatgta aaggtgttgt
     4141 aaatatgcgc aaagaggtaa tttttaatta aattaactga ataaggatta tttttaaaag
     4201 tatatgaaaa atcttgaata actcaaggtt tatttgtggt ggaatcaatt atttataata
     4261 ttgatattac catgtttatt tgtgtactct aaaaataatc agtattataa tgagcatgtt
     4321 acatctttat ttgcaggata ggaaaaccgt gttctaaaca cggtattttt atgtctattt
     4381 ttgctatata tagatatggt ataagatttt aatttgaata gcttaatcat aagttatata
     4441 attaaagttc tatattttat gtttatattt aggttatatg attatgaggc tgtaagtgaa
     4501 ttatgttcat ttacagcttt ttttgtttaa ggagattatt ttgatgattt atatatttaa
     4561 atatgaagat tatgaaaaac aaacaaattt tttaagaaat actaggatgg tgatgattaa
     4621 tggttaataa actttcagca tataaaactt atttattatt ttcagctatt acagcaatgt
     4681 gtttttcgtt agtagctaca gttatgatgg tgtatcacat tgaaatagtt catttaaatc
     4741 cacttcagct tatacttgtt ggaactactt tggaattagc atgctttata tttgaaattc
     4801 ctacagctat agttgcagat gtgtatagtc gtaaactatc tattgttatt gggggagttt
     4861 taacaggagt gggatttatt ttagaaggtt ctatttctag ttttgttttc gtacttgtag
     4921 cacagattgt atggggatta gggtctactt ttatcagtgg ctcgcttgaa gcttggattg
     4981 cggaagaaga gaagaataaa gatttagatg aaatttatat aaagggagca caagcagggc
     5041 agataggagc atttattgga atagtactaa gcactgtaat agctaatttc tctgtaaggc
     5101 ttcctattat agttagtgga gttttattta taattcttgc attattttta tggttatata
     5161 tgccagaaaa taattttaaa ccatctgctc ctggggattt aaatacattc aaaaagatgg
     5221 tatatacatt taaatctggt cttaaatttg taaaaagtaa atctataatt atgattttac
     5281 ttgcagtaac tttattttat ggattatcaa gtgaaggtta tgatagactt tctaatgcgc
     5341 attttttaca agatactaca cttcctaaac ttggaaacct tagttcagtg acttggtttg
     5401 gaatttttgg aattttagga atgatattga gcttcatagt aatgcatttt atggcaaaga
     5461 atcttaagaa tgaggataat aggaaaaatg gaaaactatt attatgcata aatatacttt
     5521 atatatcgtc tatgttgata tttgctctta caagaaactt tagtttaatg ttaatagctt
     5581 atttggcaac aaataccttt agaattataa ataaacctat attcagtgcg tggttaaatg
     5641 ggcatataga tgataattct agagctactg tgctttctat aaatggacaa atgaattcct
     5701 taggtcaaat tttaggtgga ccgattatag gaatcatagc tacaaatatt tcagtaagta
     5761 ttggtatagt atgtacttcg ttattagtaa caccggtatt agtgttatat attgttgcta
     5821 tgataattga taaaaaggtg gatgatagag ttggaggtat tgattatgaa gaaaataatt
     5881 aatataggaa tcgtagcaca cgtggatgca gtttaaacaa gatgatcttc ttgagatcgt
     5941 tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt
     6001 tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg aggagcgcag
     6061 tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa gactaactcc
     6121 tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt ccgggttgga
     6181 ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg gttcgtgcat
     6241 acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca
     6301 aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca
     6361 cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6421 actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg
     6481 gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt ccaggaaatc
     6541 tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc gtagcgagtg
     6601 ggccccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg
     6661 atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca
     6721 gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc
     6781 ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cggctaccgc
     6841 cggcgtaaca gatgagggca agcggatggc tgatgaaacc aagccaacca ggaagggcag
     6901 cccacctatc aaggtgtact gccttccaga cgaacgaaga gcgattgagg aaaaggcggc
     6961 ggcggccggc atgagcctgt cggcctacct gctggccgtc ggccagggct acaaaatcac
     7021 gggcgtcgtg gactatgagc acgtccgcga gctggcccgc atcaatggcg acctgggccg
     7081 cctgggcggc ctgctgaaac tctggctcac cgacgacccg cgcacggcgc ggttcggtga
     7141 tgccacgatc ctcgccctgc tggcgaagat cgaagagaag caggacgagc ttggcaaggt
     7201 catgatgggc gtggtccgcc cgagggcaga gccatgactt ttttagccgc taaaacggcc
     7261 ggggggtgcg cgtgattgcc aagcacgtcc ccatgcgctc catcaagaag agcgacttcg
     7321 cggagctggt gaagtacatc accgacgagc aaggcaagac cgatcgggcc c
//
