LOCUS       Exported                8573 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway recycling vector pRED56-MD8-P35S DNA, complete sequence.
ACCESSION   AB752386
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8573)
  AUTHORS   Kimura T, Shibahara K, Tanaka Y, Nakao A, Kawazu T, Nakagawa T.
  TITLE     Gateway Recycling Cloning System for Multigene Construction
  JOURNAL   Published Only in Database (2012)
REFERENCE   2  (bases 1 to 8573)
  AUTHORS   Kimura T, Shibahara K, Tanaka Y, Nakao A, Kawazu T, Nakagawa T.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-OCT-2012) Contact:Tsuyoshi Nakagawa Shimane 
            University, Center for Integrated Research in Science; 1060 
            Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE   3  (bases 1 to 8573)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     constructed using pDONR201.
FEATURES             Location/Qualifiers
     source          1..8573
                     /organism="Gateway recycling vector pRED56-MD8-P35S"
                     /mol_type="other DNA"
                     /db_xref="taxon:1237887"
     misc_feature    complement(28..127)
                     /label=attL5
                     /note="attL5"
     protein_bind    29..124
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    670..1070
                     /label=MD8
                     /note="MD8"
     regulatory      1077..1935
                     /regulatory_class="promoter"
                     /note="CaMV35S promoter"
     promoter        1582..1927
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    complement(1949..2073)
                     /label=attR1
                     /note="attR1"
     protein_bind    1949..2073
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2110..2140
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2194..2853
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="BAM62116.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            2194..2853
                     /gene="cat"
                     /label=cat
     CDS             3195..3500
                     /codon_start=1
                     /gene="ccdB"
                     /label=ccdB
                     /protein_id="BAM62117.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            3195..3500
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    3541..3665
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(3541..3665)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      3687..3951
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     terminator      3697..3949
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    complement(4158..4282)
                     /label=attR4
                     /note="attR4"
     protein_bind    4158..4282
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        4307..4337
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_feature    4609..4649
                     /label=rare cutter sites
                     /note="rare cutter sites"
     CDS             complement(4738..5598)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="BAM62118.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4738..5598)
                     /gene="bla"
                     /label=bla
     promoter        complement(5599..5703)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6000..6124
                     /label=attR3
                     /note="attR3"
     protein_bind    complement(6000..6123)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    6446..6545
                     /label=attL6
                     /note="attL6"
     protein_bind    complement(6449..6544)
                     /gene="mutant version of attL"
                     /label=attL6
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             6668..7477
                     /codon_start=1
                     /gene="kan"
                     /product="kanamycin resistance protein"
                     /label=kan
                     /protein_id="BAM62119.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            6668..7477
                     /gene="kan"
                     /label=kan
     rep_origin      7598..8237
                     /label=pUC origin
                     /note="pUC origin"
     rep_origin      7620..8208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      8343..8370
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      8462..8548
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"