LOCUS       Exported               13804 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJC24, complete sequence.
ACCESSION   KC442291
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13804)
  AUTHORS   Cheng J, Charles TC.
  TITLE     Cosmids for functional metagenomics
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13804)
  AUTHORS   Cheng J, Charles TC.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-DEC-2012) Department of Biology, University of 
            Waterloo, 200 University Avenue West, Waterloo, Ontario N2L 3G1, 
            Canada
REFERENCE   3  (bases 1 to 13804)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13804
                     /organism="Cloning vector pJC24"
                     /mol_type="other DNA"
                     /db_xref="taxon:1294007"
     primer_bind     252..268
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     276..381
                     /label=attL
                     /note="attL"
     protein_bind    282..381
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(402..1823)
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /protein_id="AGF38328.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(402..1823)
                     /gene="sacB"
                     /label=sacB
     misc_recomb     2206..2305
                     /label=attL
                     /note="attL"
     protein_bind    complement(2206..2305)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2324..2340)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2348..2364
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2372..2402)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2417..2438
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(2670..3041)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /protein_id="AGF38329.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     gene            complement(2670..3041)
                     /gene="traJ"
                     /label=traJ
     oriT            3074..3183
                     /note="oriT"
     CDS             complement(3864..4514)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=tetR
                     /protein_id="AGF38330.1"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     gene            complement(3864..4514)
                     /gene="tetR"
                     /label=tetR
     CDS             4620..5819
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline resistance protein"
                     /label=tetA
                     /protein_id="AGF38331.1"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     gene            4620..5819
                     /gene="tetA"
                     /label=tetA
     misc_recomb     6187..6419
                     /label=lambda cos site
                     /note="lambda cos site"
     CDS             complement(8218..9366)
                     /codon_start=1
                     /gene="trfA"
                     /product="trans-acting replication protein"
                     /label=trfA
                     /protein_id="AGF38332.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(8218..9366)
                     /gene="trfA"
                     /label=trfA
     CDS             complement(9415..9765)
                     /codon_start=1
                     /gene="ssB"
                     /product="single-stranded DNA binding protein"
                     /label=ssB
                     /protein_id="AGF38333.1"
                     /translation="MSHNQFQFIGNLTRDTEVRHGNSNKPQAIFDIAVNEEWRNDAGDK
                     QERTDFFRIKCFGSQAEAHGKYLGKGSLVFVQGKIRNTKYEKDGQTVYGTDFIADKVDY
                     LDTKAPGGSNQE"
     gene            complement(9415..9765)
                     /gene="ssB"
                     /label=ssB
     CDS             9940..10251
                     /codon_start=1
                     /gene="trbA"
                     /product="TrbA"
                     /label=trbA
                     /note="regulatory protein"
                     /protein_id="AGF38334.1"
                     /translation="MTKHELSERAGVSISFLSDLTNGKANPSLKVMEAIADALETPLPL
                     LLESTDLDREALAEIAGHPFKSSVPPGYERISVVLPSHKAFIVKKWGDDTRKKLRGRL"
     gene            9940..10251
                     /gene="trbA"
                     /label=trbA
     CDS             complement(11289..12383)
                     /codon_start=1
                     /gene="incC"
                     /product="plasmid maintenance protein"
                     /label=incC
                     /protein_id="AGF38336.1"
                     /translation="MGVIHEETAYRKPVPGGDPGAGSGAADHRDSAGRLSRWEATGDVR
                     NVAGTDQGRSVASGASRVGRVRGQELARGVRAGNGGSAGTSGVHRPEVGSGRQEKTGNQ
                     TMKTLVTANQKGGVGKTSTLVHLAFDFFERGLRVAVIDLDPQGNASYTLKDFATGLHAS
                     KLFGAVPAGGWTETAPAAGDGQAARLALIESNPVLANAERLSLDDARELFGANIKALAN
                     QGFDVCLIDTAPTLGVGLAAALFAADYVLSPIELEAYSIQGIKKMVTTIANVRQKNAKL
                     QFLGMVPSKVDARNPRHARHQAELLAAYPKMMIPATVGLRSSIADALASGVPVWKIKKT
                     AARKASKEVRALADYVFTKMEISQ"
     gene            complement(11289..12383)
                     /gene="incC"
                     /label=incC
     CDS             complement(12065..12370)
                     /codon_start=1
                     /gene="korA"
                     /product="KorA"
                     /label=korA
                     /note="regulatory protein"
                     /protein_id="AGF38335.1"
                     /translation="MKKRLTESQFQEAIQGLEVGQQTIEIARGVLVDGKPQATFATSLG
                     LTRGAVSQAVHRVWAAFEDKNLPEGYARVTAVLPEHQAYIVRKWEADAKKKQETKR"
     gene            complement(12065..12370)
                     /gene="korA"
                     /label=korA
     rep_origin      12835..13546
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      12845..13539
                     /label=ori (RK2)
                     /note="ori (RK2)"