LOCUS       40924_36508        8120 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway recycling vector pRED56-MD8-19 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8120)
  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  JOURNAL   Unpublished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 8120)
  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 8120)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8120)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-NOV-2015) Contact:Tsuyoshi Nakagawa Shimane University, 
            Interdisciplinary Center for Science Research; 1060 Nishikawatsu, 
            Matsue, Shimane 690-8504, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     constructed using pDONR201.
FEATURES             Location/Qualifiers
     source          1..8120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    28..127
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    674..1071
                     /label=MD8
                     /note="MD8"
     protein_bind    1094..1218
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1255..1285
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1339..1995
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2340..2642
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2686..2810)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2836..2865
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             2872..2901
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2917..2946
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2953..2982
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2989..3018
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3034..3063
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3070..3099
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3106..3135
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3151..3180
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3187..3216
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      3244..3496
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    3705..3829
                     /label=attR4
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3854..3884
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_feature    4156..4196
                     /label=rare cutter sites
                     /note="rare cutter sites"
     CDS             complement(4288..5145)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5146..5250)
                     /label=AmpR promoter
     protein_bind    complement(5547..5670)
                     /label=attR3
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(5996..6091)
                     /label=attL6
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             6215..7021
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      7167..7755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7890..7917)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(8009..8095)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 gttaacgcta gcatggatct cgggccccaa ataatgattt tattttgact gatagtgacc
       61 tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt gtatacaaaa
      121 gtaggctgga tgaatcgaat aacttcttct tcaaatgcgc aaacgaagaa atttcctcag
      181 tctcagccac agccgccgga gaaaaatgag aaaaaactga aaagggagag atttccggtg
      241 aaccctttga gagatttcag tacaagaagc tctggaagca gcagttgtag caatgagagt
      301 gcttctggtt caacttccgg tgaagcatcc aacggttgtc acaggttctt gctctctcat
      361 tctttctcat cgtcttcttc atcatcttct tcttcgttgg gagtttttcc tcgtagacca
      421 gtgagatcag tagctaaaaa ccccaaatct gctccggtag ttagtaaacc cttaattaga
      481 aagaaaccaa gctctcttga ggaggttaag ttgaaatcta ctcttactga gaaacctaat
      541 ttgctgaagt ctcagagatg caagacaaat cctgtatctg gaaagagacc aacttgtaag
      601 attaccatga aaccggagaa agtttcggtt ttgaagaaac aatcttctgt ttctaggtaa
      661 gctgcagtgt tttcatcaca cgtaatatca tccattttat attatgatcc acaaacagca
      721 aaggcccatt aatctcatta tattttagat taattgattg caattgccca acttctccac
      781 tcagcaatgt tagttttaaa gacacatcaa gttagtgtat tttattttta atctttctat
      841 aaaaaacaac actttgtttt tgtctatttc ttattctgtg aaaaagagaa atgaaaaagt
      901 aaaaaagata acaaaaaggc tggtttttca cggtgagaat ttttcatggt cacgcaataa
      961 cttaaatatg aactattaaa ataactaaga ttgaaaaatt agcatgatca aaagaagaag
     1021 aaaaaaaaga caacatgaag cggaaaggaa aaaaaagaaa gctgaaaatc taagcttggg
     1081 tctagagtta tcaacaagtt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa
     1141 tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca
     1201 acatatccag tcactatggc ggcccctcga ggggccgcat taggcacccc aggctttaca
     1261 ctttatgctt ccggctcgta taatgtgtgg attttgagtt aggatccggc gagattttca
     1321 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     1381 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     1441 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     1501 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt
     1561 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     1621 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     1681 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     1741 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
     1801 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     1861 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     1921 gtctgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     1981 tggcagggcg gggcgtaaag atctggatcc ggcttactaa aagccagata acagtatgcg
     2041 tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg
     2101 tcaaaaagag gtgtgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc
     2161 agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa
     2221 tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga
     2281 ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg actggtgaaa
     2341 tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     2401 agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc
     2461 tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     2521 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
     2581 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     2641 tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag tgactggata
     2701 tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg
     2761 atatttatat cattttacgt ttctcgttca gctttcttgt acaaagtggt tgataacagc
     2821 gggttaatta acggtgaaca aaagctaatc tccgaggaag acttgaacgg tgaacaaaaa
     2881 ttaatctcag aagaagactt gaacggactc gacggtgaac aaaagttgat ttctgaagaa
     2941 gatttgaacg gtgaacaaaa gctaatctcc gaggaagact tgaacggtga acaaaaatta
     3001 atctcagaag aagacttgaa cggactcgac ggtgaacaaa agttgatttc tgaagaagat
     3061 ttgaacggtg aacaaaagct aatctccgag gaagacttga acggtgaaca aaaattaatc
     3121 tcagaagaag acttgaacgg actcgacggt gaacaaaagt tgatttctga agaagatttg
     3181 aacggtgaac aaaagctaat ctccgaggaa gacttgaacg gtgcttagag ctcgaatttc
     3241 cccgatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt
     3301 gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa
     3361 tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa
     3421 tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca
     3481 tctatgttac tagatcggga attctagctt cgacaattat tttggactct catgttctca
     3541 agctgaaatc caaaagaaac gtgatgttgg ttcagatttg gaatcactga caatgaagtt
     3601 tcagtcagcc ggtttgtcac caaggatcca ggcttcatca tgggagccga gccgttcaag
     3661 tttcaacttc gactatttgg atccgtccta gtggatcccc catcacaact ttgtatagaa
     3721 aagttgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
     3781 aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg cggccgcatt
     3841 aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga ttttgagtta
     3901 ggatccggcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
     3961 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     4021 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     4081 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     4141 tgctcatccg gaattgcgat cgcatttaaa tggccggccc ttaattaagg cgcgccgttt
     4201 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4261 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4321 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4381 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4441 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4501 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4561 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg
     4621 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4681 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4741 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4801 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4861 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     4921 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4981 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5041 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5101 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5161 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5221 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaac
     5281 gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt
     5341 gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg
     5401 gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat
     5461 gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata
     5521 cacagccagt ctgcaggtcg atcgaccata gtgactggat atgttgtgtt ttacagtatt
     5581 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     5641 tttctcgttc aactttatta tacaaagttg tgatccacat cttctgattc tattgatctc
     5701 tcacagtttc agagaggttt agttgacagg agttcttgtc atcctcatgt aacactggat
     5761 aaagtgtcaa ggacacattt gagagtagag caaacaaata gccacatgcc tgaaatgaaa
     5821 tcccaacaga ttacagatac tgaatttgat attcaaaatc acaaagaaag tgctgcttct
     5881 ctgggaaaag gaagagaatt gcttccatgc tctgctgcag agtcaataag cactgatgga
     5941 ggtggtctga tttgttctga ggattcaaat tggatggcat gttacaagaa ttacccaact
     6001 ttttaataca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     6061 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgcagctct ggcccgtgtc
     6121 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     6181 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     6241 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     6301 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     6361 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     6421 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     6481 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     6541 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     6601 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     6661 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     6721 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     6781 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     6841 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     6901 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     6961 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     7021 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     7081 cggcgcaagc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7141 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7201 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7261 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     7321 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7381 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7441 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7501 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7561 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7621 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7681 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7741 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7801 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7861 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     7921 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     7981 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     8041 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     8101 tggcagttcc ctactctcgc
//
