LOCUS       Exported                7922 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Clostridium difficile shuttle vector pRAN334, complete sequence.
ACCESSION   KT371994
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7922)
  AUTHORS   Ransom EM, Williams KB, Weiss DS, Ellermeier CD.
  TITLE     Identification and characterization of a gene cluster required for 
            proper rod shape, cell division, and pathogenesis in Clostridium 
            difficile
  JOURNAL   J. Bacteriol. 196 (12), 2290-2300 (2014)
  PUBMED    24727226
REFERENCE   2  (bases 1 to 7922)
  AUTHORS   Ransom EM, Weiss DS, Ellermeier CD.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-AUG-2015) Microbiology, The University of Iowa, 51 
            Newton Rd, Iowa City, IA 52242, USA
REFERENCE   3  (bases 1 to 7922)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7922
                     /organism="Clostridium difficile shuttle vector pRAN334"
                     /mol_type="other DNA"
                     /db_xref="taxon:1780072"
     CDS             complement(55..681)
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /label=tetR
                     /protein_id="AMA07789.1"
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     gene            complement(55..681)
                     /gene="tetR"
                     /label=tetR
     CDS             complement(55..678)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     protein_bind    788..806
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             873..1592
                     /codon_start=1
                     /gene="cfpOpt"
                     /product="CFPopt"
                     /label=cfpOpt
                     /protein_id="AMA07790.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            873..1592
                     /gene="cfpOpt"
                     /label=cfpOpt
     CDS             complement(1938..2309)
                     /codon_start=1
                     /gene="traJ"
                     /product="TraJ"
                     /label=traJ
                     /protein_id="AMA07793.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     gene            complement(1938..2309)
                     /gene="traJ"
                     /label=traJ
     oriT            2342..2451
                     /note="incP origin of transfer"
     CDS             complement(2827..3450)
                     /codon_start=1
                     /gene="catP"
                     /product="CatP"
                     /label=catP
                     /protein_id="AMA07791.1"
                     /translation="MVFEKIDKNSWNRKEYFDHYFASVPCTYSMTVKVDITQIKEKGMK
                     LYPAMLYYIAMIVNRHSEFRTAINQDGELGIYDEMIPSYTIFHNDTETFSSLWTECKSD
                     FKSFLADYESDTQRYGNNHRMEGKPNAPENIFNVSMIPWSTFDGFNLNLQKGYDYLIPI
                     FTMGKYYKEDNKIILPLAIQVHHAVCDGFHICRFVNELQELINS"
     gene            complement(2827..3450)
                     /gene="catP"
                     /label=catP
     rep_origin      complement(3798..4417)
                     /direction=LEFT
     rep_origin      complement(3813..4401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5194..6831)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="plasmid replication; derived from Clostridium 
                     difficile"
                     /protein_id="AMA07794.1"
                     /translation="MEQLDSKYKLKKFLMAVFRDGIGQGNNLIDNEYVRVFQNNKSNSK
                     QLELGEEFKEYSKTTFFKNIDDIVEFTFAKNIYYENTFFNLCTTDGKAGTNENLINRYA
                     LGFDFDKKELGQGFNYKDIINLFTKIGLHYHILVDSGNGFHVYVLINKTNNIKLVSEVT
                     NTLINKLGADKQANLSTQVLRVPYTYNIKNTTKQVKIIHQDKNIYRYDIEKLAKKYCKD
                     VKTVGNTNTKYILDSKLPNCIVDILKNGSKDGHKNLDLQKIVVTLRLRNKSLSQVISVA
                     REWNYISQNSLSNSELEYQVKYMYEKLKTVNFGCTGCEFNSDCWNKIESDFIYSDEDTL
                     FNMPHKHSKDLKYKNRKGVKIMTGNQLFIYNVLLNNKDRELNIDDIMELITYKRKKKVK
                     NIVMSEKTLRETLKELQHNDYITKTKGVTKLGIKDTYNVKEVRCNIDKQYTISYFVTMA
                     VIWGIISTEELRLYTHMRYKQDLLVKDDKIKGNILRINQEELAKDLGVTQQRISNMIES
                     LLDTKILDVWETKINDRGFMYYTYRLNK"
     gene            complement(5194..6831)
                     /gene="repA"
                     /label=repA
     CDS             complement(7106..7618)
                     /codon_start=1
                     /gene="orfB"
                     /product="OrfB"
                     /label=orfB
                     /protein_id="AMA07792.1"
                     /translation="MRARSSKWRYHIRTNNKSICCNIKNFIHNLELFYKMELKLSDNII
                     NDKLYYSNIAEFEEFETLEKAREVESTIISQYQFLDSINHMLKQKIILLSNKDSVLNIT
                     KNGNTNYLKVKNKYIEKHKNKPIMRYHINCQFNTDGSVKSITQEFEPILELNKKNTLSR
                     PSRVFLK"
     gene            complement(7106..7618)
                     /gene="orfB"
                     /label=orfB