LOCUS       Exported                5005 bp ds-DNA    circular SYN 09-11-2013
DEFINITION  Donor vector for inserting an attB-flanked gene to generate an entry
            vector in the Gateway(R) system.
ACCESSION   .
VERSION     .
KEYWORDS    pDONR223
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5005)
  AUTHORS   Rual JF, Hirozane-Kishikawa T, Hao T, Bertin N, Li S, Dricot A, Li 
            N, Rosenberg J, Lamesch P, Vidalain PO, Clingingsmith TR, Hartley 
            JL, Esposito D, Cheo D, Moore T, Simmons B, Sequerra R, Bosak S, 
            Doucette-Stamm L, Le Peuch C, Vandenhaute J, Cusick ME, Albala JS, 
            Hill DE, Vidal M.
  TITLE     Human ORFeome version 1.1: a platform for reverse proteomics.
  JOURNAL   Genome Res. 2004;14:2128-35.
  PUBMED    15489335
REFERENCE   2  (bases 1 to 5005)
  AUTHORS   I.M.A.G.E. Consortium
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5005
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     terminator      268..295
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     537..553
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    570..801
                     /gene="mutant version of attP"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attP1"
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)221 version)"
     CDS             complement(1197..1502)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /note="ccdB"
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagaged in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(1844..2503)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(2504..2606)
                     /note="cat promoter"
                     /note="promoter of the E. coli cat gene"
     protein_bind    complement(2751..2982)
                     /gene="mutant version of attP"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attP2"
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)221 version)"
     promoter        complement(3001..3019)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3024..3040)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3471..4262
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase"
                     /note="SmR"
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      4355..4943
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"