LOCUS       Exported                7371 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMTL84422, complete sequence.
ACCESSION   FJ797650
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7371
                     /organism="Shuttle vector pMTL84422"
                     /mol_type="other DNA"
                     /db_xref="taxon:647633"
     regulatory      9..62
                     /regulatory_class="terminator"
                     /note="terminator derived from CD0164 gene of Clostridium 
                     difficile"
     primer_bind     63..80
                     /label=M13R
                     /note="M13R"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      89..238
                     /regulatory_class="promoter"
                     /note="promoter derived from thl gene of Clostridium 
                     acetobutylicum"
     misc_feature    242..505
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             242..505
                     /codon_start=1
                     /gene="lacZ alpha"
                     /product="beta-galactosidase, alpha subunit"
                     /label=lacZ alpha
                     /protein_id="ACR43912.1"
                     /translation="MTMITNSSSVPGDPLESTSRVHGDLEACRHASLALAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     gene            242..505
                     /gene="lacZ alpha"
                     /label=lacZ alpha
     primer_bind     complement(343..360)
                     /label=M13F
                     /note="M13F"
     primer_bind     complement(343..359)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      507..548
                     /regulatory_class="terminator"
                     /note="terminator derived from fdx gene of Clostridium 
                     pasteurianum"
     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      complement(5919..6480)
                     /direction=LEFT
                     /label=p15A
                     /note="p15A"
     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
                     /note="incP origin of transfer"
     CDS             6797..7237
                     /codon_start=1
                     /gene="traJ"
                     /product="plasmid conjugative transfer protein"
                     /label=traJ
                     /protein_id="ACR43911.1"
                     /translation="MLNGNPALRGWPATAGVTDEGKRMADETKPTRKGSPPIKVYCLPD
                     ERRAIEEKAAAAGMSLSAYLLAVGQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWL
                     TDDPRTARFGDATILALLAKIEEKQDELGKVMMGVVRPRAEP"
     gene            6797..7237
                     /gene="traJ"
                     /label=traJ
     CDS             6866..7237
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"