LOCUS       Exported                8038 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway recycling vector pRED56-MD8-Pnos-09 DNA, complete sequence.
ACCESSION   LC094277
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8038)
  AUTHORS   Saisho T, Shibahara K, Nakao A, Tanaka Y, Nishimura K, Kimura T, 
            Nakagawa T.
  TITLE     Improved Gateway Recycling Cloning System for Multigene Construction
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8038)
  AUTHORS   Saisho T, Shibahara K, Nakao A, Tanaka Y, Nishimura K, Kimura T, 
            Nakagawa T.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-NOV-2015) Contact:Tsuyoshi Nakagawa Shimane 
            University, Interdisciplinary Center for Science Research; 1060 
            Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE   3  (bases 1 to 8038)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     constructed using pDONR201.
FEATURES             Location/Qualifiers
     source          1..8038
                     /organism="Gateway recycling vector pRED56-MD8-Pnos-09"
                     /mol_type="other DNA"
                     /db_xref="taxon:1747838"
     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    674..1071
                     /label=MD8
                     /note="MD8"
     regulatory      1078..1380
                     /regulatory_class="promoter"
                     /note="NOS promoter"
     promoter        1160..1339
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        1192..1375
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    1393..1410
                     /label=6xHis
                     /note="6xHis"
     CDS             1393..1410
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    complement(1420..1544)
                     /label=attR1
                     /note="attR1"
     protein_bind    1420..1544
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1581..1611
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1665..2324
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="BAT33378.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            1665..2324
                     /gene="cat"
                     /label=cat
     CDS             2666..2971
                     /codon_start=1
                     /gene="ccdB"
                     /label=ccdB
                     /protein_id="BAT33379.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            2666..2971
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    3012..3136
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(3012..3136)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      3152..3416
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     terminator      3162..3414
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    complement(3623..3747)
                     /label=attR4
                     /note="attR4"
     protein_bind    3623..3747
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3772..3802
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_feature    4074..4114
                     /label=rare cutter sites
                     /note="rare cutter sites"
     CDS             complement(4203..5063)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="BAT33380.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4203..5063)
                     /gene="bla"
                     /label=bla
     promoter        complement(5064..5168)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5465..5589
                     /label=attR3
                     /note="attR3"
     protein_bind    complement(5465..5588)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    5911..6010
                     /label=attL6
                     /note="attL6"
     protein_bind    complement(5914..6009)
                     /gene="mutant version of attL"
                     /label=attL6
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             6133..6942
                     /codon_start=1
                     /gene="kan"
                     /product="kanamycin resistance protein"
                     /label=kan
                     /protein_id="BAT33381.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            6133..6942
                     /gene="kan"
                     /label=kan
     rep_origin      7063..7702
                     /label=pUC origin
                     /note="pUC origin"
     rep_origin      7085..7673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      7808..7835
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      7927..8013
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"