LOCUS       Exported               13053 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJC8, complete sequence.
ACCESSION   KC149513
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13053)
  AUTHORS   Lam KN, Hall MW, Engel K, Vey G, Cheng J, Neufeld JD, Charles TC.
  TITLE     Evaluation of a pooled strategy for high-throughput sequencing of 
            cosmid clones from metagenomic libraries
  JOURNAL   PLoS ONE 9 (6), E98968 (2014)
  PUBMED    24911009
REFERENCE   2  (bases 1 to 13053)
  AUTHORS   Cheng J, Charles TC.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-NOV-2012) Biology, University of Waterloo, 200 
            University Avenue West, Waterloo, Ontario N2L 3G1, Canada
REFERENCE   3  (bases 1 to 13053)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13053
                     /organism="Cloning vector pJC8"
                     /mol_type="other DNA"
                     /db_xref="taxon:1273179"
     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(450..1235)
                     /codon_start=1
                     /gene="accC4"
                     /product="gentamicin resistance"
                     /label=accC4
                     /protein_id="AGA63621.1"
                     /translation="MQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRPLEDGPLGLI
                     EALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNVKRSAHPFAF
                     AAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLHLAELMAKVP
                     YGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGHAFARLIRSR
                     DIVATALGQLGRDPLIFLHPPEGGMRRMRCRSPVDWLSS"
     gene            complement(450..1235)
                     /gene="accC4"
                     /label=accC4
     misc_recomb     1455..1554
                     /label=attL
                     /note="attL"
     protein_bind    complement(1455..1554)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1573..1589)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1597..1613
                     /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(1621..1651)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1666..1687
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(1919..2290)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /note="conjugation"
                     /protein_id="AGA63622.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     gene            complement(1919..2290)
                     /gene="traJ"
                     /label=traJ
     oriT            2323..2432
                     /label=incP origin of transfer
                     /note="incP origin of transfer"
     CDS             complement(3113..3763)
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /label=tetR
                     /note="tetracycline resistance regulation"
                     /protein_id="AGA63623.1"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     gene            complement(3113..3763)
                     /gene="tetR"
                     /label=tetR
     CDS             3869..5068
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline resistance"
                     /label=tetA
                     /protein_id="AGA63625.1"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     gene            3869..5068
                     /gene="tetA"
                     /label=tetA
     misc_recomb     5436..5668
                     /note="cos; lambda cos site"
     CDS             complement(7467..8615)
                     /codon_start=1
                     /gene="trfA"
                     /product="trans-acting replication"
                     /label=trfA
                     /protein_id="AGA63626.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(7467..8615)
                     /gene="trfA"
                     /label=trfA
     CDS             complement(8664..9014)
                     /codon_start=1
                     /gene="ssB"
                     /product="single-stranded DNA binding protein"
                     /label=ssB
                     /protein_id="AGA63627.1"
                     /translation="MSHNQFQFIGNLTRDTEVRHGNSNKPQAIFDIAVNEEWRNDAGDK
                     QERTDFFRIKCFGSQAEAHGKYLGKGSLVFVQGKIRNTKYEKDGQTVYGTDFIADKVDY
                     LDTKAPGGSNQE"
     gene            complement(8664..9014)
                     /gene="ssB"
                     /label=ssB
     CDS             9189..9500
                     /codon_start=1
                     /gene="trbA"
                     /product="TrbA"
                     /label=trbA
                     /note="regulation of tra gene expression"
                     /protein_id="AGA63628.1"
                     /translation="MTKHELSERAGVSISFLSDLTNGKANPSLKVMEAIADALETPLPL
                     LLESTDLDREALAEIAGHPFKSSVPPGYERISVVLPSHKAFIVKKWGDDTRKKLRGRL"
     gene            9189..9500
                     /gene="trbA"
                     /label=trbA
     CDS             complement(10538..11632)
                     /codon_start=1
                     /gene="incC"
                     /product="plasmid maintenance protein"
                     /label=incC
                     /protein_id="AGA63629.1"
                     /translation="MGVIHEETAYRKPVPGGDPGAGSGAADHRDSAGRLSRWEATGDVR
                     NVAGTDQGRSVASGASRVGRVRGQELARGVRAGNGGSAGTSGVHRPEVGSGRQEKTGNQ
                     TMKTLVTANQKGGVGKTSTLVHLAFDFFERGLRVAVIDLDPQGNASYTLKDFATGLHAS
                     KLFGAVPAGGWTETAPAAGDGQAARLALIESNPVLANAERLSLDDARELFGANIKALAN
                     QGFDVCLIDTAPTLGVGLAAALFAADYVLSPIELEAYSIQGIKKMVTTIANVRQKNAKL
                     QFLGMVPSKVDARNPRHARHQAELLAAYPKMMIPATVGLRSSIADALASGVPVWKIKKT
                     AARKASKEVRALADYVFTKMEISQ"
     gene            complement(10538..11632)
                     /gene="incC"
                     /label=incC
     CDS             complement(11314..11619)
                     /codon_start=1
                     /gene="korA"
                     /product="KorA"
                     /label=korA
                     /note="regulation of gene expression"
                     /protein_id="AGA63624.1"
                     /translation="MKKRLTESQFQEAIQGLEVGQQTIEIARGVLVDGKPQATFATSLG
                     LTRGAVSQAVHRVWAAFEDKNLPEGYARVTAVLPEHQAYIVRKWEADAKKKQETKR"
     gene            complement(11314..11619)
                     /gene="korA"
                     /label=korA
     rep_origin      12084..12795
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      12094..12788
                     /note="oriV; incP origin of replication; RK2"