LOCUS       Exported                7548 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCIE, complete sequence.
ACCESSION   KY774666
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7548)
  AUTHORS   Weaver KE, Chen Y, Miiller EM, Johnson JN, Dangler AA, Manias DA, 
            Clem AM, Schjodt DJ, Dunny GM.
  TITLE     Examination of E. faecalis toxin-antitoxin (TA) toxin Fst function 
            utilizing a pheromone-inducible expression vector with tight 
            repression and large dynamic range
  JOURNAL   J. Bacteriol. (2017) In press
  PUBMED    28348028
REFERENCE   2  (bases 1 to 7548)
  AUTHORS   Weaver K, Chen Y, Miiller E, Johnson J, Dangler A, Manias D, Clem A,
            Schjodt D, Dunny G.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-MAR-2017) Department of Microbiology & Immunology, 
            University of Minnesota, 689 23rd Avenue S.E., Minneapolis, MN 
            55455, USA
REFERENCE   3  (bases 1 to 7548)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7548
                     /organism="Cloning vector pCIE"
                     /mol_type="other DNA"
                     /note="derived from Escherichia coli and Enterococcus 
                     faecalis"
                     /db_xref="taxon:1977276"
     CDS             complement(74..1027)
                     /codon_start=1
                     /gene="prgX"
                     /product="PrgX"
                     /label=prgX
                     /protein_id="ARF06186.1"
                     /translation="MFKIGSVLKQIRQELNYHQIDLYSGIMSKSVYIKVEADSRPISVE
                     ELSKFSERLGVNFFEILNRAGMNTKSVNETGKEKLLISKIFTNPDLFDKNFQRIEPKRL
                     TSLQYFSIYLGYISIAHHYNIEVPTFNKTITSDLKHLYDKRTTFFGIDYEIVSNLLNVL
                     PYEEVSSIIKPMYPIVDSFGKDYDLTIQTVLKNALTISIMNRNLKEAQYYINQFEHLKT
                     IKNISINGYYDLEINYLKQIYQFLTDKNIDSYLNAVNIINIFKIIGKEDIHRSLVEELT
                     KISAKEKFTPPKEVTMYYENYVAIENNPIPEIKEQS"
     gene            complement(74..1027)
                     /gene="prgX"
                     /label=prgX
     regulatory      1206..1214
                     /regulatory_class="promoter"
     CDS             1236..1307
                     /codon_start=1
                     /gene="prgQ"
                     /product="PrgQ"
                     /label=prgQ
                     /protein_id="ARF06187.1"
                     /translation="MKTTLKKLSRYIAVVIAITLIFI"
     gene            1236..1307
                     /gene="prgQ"
                     /label=prgQ
     regulatory      1556..1600
                     /regulatory_class="terminator"
     regulatory      1699..1749
                     /regulatory_class="terminator"
     CDS             complement(2152..2163)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2625..3275
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="ARF06188.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            2625..3275
                     /gene="cat"
                     /label=cat
     CDS             complement(3619..4779)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B"
                     /label=repB
                     /protein_id="ARF06189.1"
                     /translation="MSSISKNEPNQKQVQTLNELSKRKVVEHNSLITSIAKMDKTPLKM
                     FELAVSCINTEEPPKDHTVYLSKEELFAFFKVSDNDKHSRFKQAVENMQKQAFFQIKEE
                     VGKGFKFRSIVPIPYVEWTDYHDDVKIEFHREIMPYLINLKQNFTQHALSDIAELNSKY
                     SIILYRWLSMNYNQYEHYSYKGGRREEQVEAYRNPTISMRELREMTDTVDEYPRFDRLE
                     HRVLKEPIEEINENTSFNVTYDKIKKGRSIDSIVFHITKKRRADDNSYKLEDKDYQSDK
                     EEKSRNEADLLKQAMESKYTRLLIENFLLSPLEMTDTALMAGLQKNVYPLYDELKELRG
                     LNGVKDHLSYISSKKEAYSKRNVAKYLKKAIEQYLPTVKRQDLNHE"
     gene            complement(3619..4779)
                     /gene="repB"
                     /label=repB
     misc_feature    6144..6284
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(6470..7058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"