LOCUS       Exported                7952 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Clostridium difficile shuttle vector pRAN473, complete sequence.
ACCESSION   KT371996
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7952)
  AUTHORS   Ransom EM, Ellermeier CD, Weiss DS.
  TITLE     Use of mCherry Red fluorescent protein for studies of protein 
            localization and gene expression in Clostridium difficile
  JOURNAL   Appl. Environ. Microbiol. 81 (5), 1652-1660 (2015)
  PUBMED    25527559
REFERENCE   2  (bases 1 to 7952)
  AUTHORS   Ransom EM, Ellermeier CD, Weiss DS.
  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 7952)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7952
                     /organism="Clostridium difficile shuttle vector pRAN473"
                     /mol_type="other DNA"
                     /db_xref="taxon:1780075"
     CDS             complement(55..681)
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /label=tetR
                     /protein_id="AMA07804.1"
                     /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..1640
                     /codon_start=1
                     /gene="mCherryOpt-MCS"
                     /product="mCherryOpt-MCS"
                     /label=mCherryOpt-MCS
                     /protein_id="AMA07805.1"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGDFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYKMNACVDLIKQLRRAGSYKF"
     gene            873..1640
                     /gene="mCherryOpt-MCS"
                     /label=mCherryOpt-MCS
     CDS             complement(1968..2339)
                     /codon_start=1
                     /gene="traJ"
                     /product="TraJ"
                     /label=traJ
                     /protein_id="AMA07806.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     gene            complement(1968..2339)
                     /gene="traJ"
                     /label=traJ
     oriT            2372..2481
                     /note="incP origin of transfer"
     CDS             complement(2857..3480)
                     /codon_start=1
                     /gene="catP"
                     /product="CatP"
                     /label=catP
                     /protein_id="AMA07803.1"
                     /translation="MVFEKIDKNSWNRKEYFDHYFASVPCTYSMTVKVDITQIKEKGMK
                     LYPAMLYYIAMIVNRHSEFRTAINQDGELGIYDEMIPSYTIFHNDTETFSSLWTECKSD
                     FKSFLADYESDTQRYGNNHRMEGKPNAPENIFNVSMIPWSTFDGFNLNLQKGYDYLIPI
                     FTMGKYYKEDNKIILPLAIQVHHAVCDGFHICRFVNELQELINS"
     gene            complement(2857..3480)
                     /gene="catP"
                     /label=catP
     rep_origin      3828..4447
                     /label=colE1
                     /note="colE1"
     rep_origin      complement(3843..4431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5224..6861)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="plasmid replication; derived from Clostridium 
                     difficile"
                     /protein_id="AMA07802.1"
                     /translation="MEQLDSKYKLKKFLMAVFRDGIGQGNNLIDNEYVRVFQNNKSNSK
                     QLELGEEFKEYSKTTFFKNIDDIVEFTFAKNIYYENTFFNLCTTDGKAGTNENLINRYA
                     LGFDFDKKELGQGFNYKDIINLFTKIGLHYHILVDSGNGFHVYVLINKTNNIKLVSEVT
                     NTLINKLGADKQANLSTQVLRVPYTYNIKNTTKQVKIIHQDKNIYRYDIEKLAKKYCKD
                     VKTVGNTNTKYILDSKLPNCIVDILKNGSKDGHKNLDLQKIVVTLRLRNKSLSQVISVA
                     REWNYISQNSLSNSELEYQVKYMYEKLKTVNFGCTGCEFNSDCWNKIESDFIYSDEDTL
                     FNMPHKHSKDLKYKNRKGVKIMTGNQLFIYNVLLNNKDRELNIDDIMELITYKRKKKVK
                     NIVMSEKTLRETLKELQHNDYITKTKGVTKLGIKDTYNVKEVRCNIDKQYTISYFVTMA
                     VIWGIISTEELRLYTHMRYKQDLLVKDDKIKGNILRINQEELAKDLGVTQQRISNMIES
                     LLDTKILDVWETKINDRGFMYYTYRLNK"
     gene            complement(5224..6861)
                     /gene="repA"
                     /label=repA
     CDS             complement(7136..7648)
                     /codon_start=1
                     /gene="OrfB"
                     /product="orfB"
                     /label=OrfB
                     /protein_id="AMA07801.1"
                     /translation="MRARSSKWRYHIRTNNKSICCNIKNFIHNLELFYKMELKLSDNII
                     NDKLYYSNIAEFEEFETLEKAREVESTIISQYQFLDSINHMLKQKIILLSNKDSVLNIT
                     KNGNTNYLKVKNKYIEKHKNKPIMRYHINCQFNTDGSVKSITQEFEPILELNKKNTLSR
                     PSRVFLK"
     gene            complement(7136..7648)
                     /gene="OrfB"
                     /label=OrfB