LOCUS       40924_36463        5401 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway recycling vector pRED418-Amp DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5401)
  AUTHORS   Kimura T, Nakao A, Murata S, Kobayashi Y, Tanaka Y, Shibahara K, 
            Kawazu T, Nakagawa T.
  TITLE     Development of the gateway recycling cloning system for multiple 
            linking of expression cassettes in a defined order, and direction on
            gateway compatible binary vectors
  JOURNAL   Biosci. Biotechnol. Biochem. 77 (2), 430-434 (2013)
  PUBMED    23391940
REFERENCE   2  (bases 1 to 5401)
  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 5401)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5401)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biosci. 
            Biotechnol. Biochem."; date: "2013"; volume: "77"; issue: "2"; 
            pages: "430-434"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-OCT-2012) Contact:Tsuyoshi Nakagawa Shimane University, Center 
            for Integrated Research in Science; 1060 Nishikawatsu, Matsue, 
            Shimane 690-8504, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     constructed using pDONR201.
FEATURES             Location/Qualifiers
     source          1..5401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    28..127
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     179..195
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    196..252
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(265..281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    803..927
                     /label=attR4
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        952..982
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     misc_feature    1254..1294
                     /label=rare cutter sites
                     /note="rare cutter sites"
     CDS             complement(1386..2243)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2244..2348)
                     /label=AmpR promoter
     protein_bind    complement(2645..2768)
                     /label=attR3
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3274..3373)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3496..4302
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      4448..5036
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5171..5198)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(5290..5376)
                     /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 taagcaatgc ttttttataa tgccaacttt gtacaaaaaa
      121 gcaggctgga tgaatcgaat aacttcttct tcaaatgcgt tttcccagtc acgacgttgt
      181 aaaacgacgg ccagtgaatt cgagctcggt acccggggat cctctagagt cgacctgcag
      241 gcatgcaagc ttggcgtaat catggtcata gctgtttcct ggcaaacgaa gaaatttcct
      301 cagtctcagc cacagccgcc ggagaaaaat gagaaaaaac tgaaaaggga gagatttccg
      361 gtgaaccctt tgagagattt cagtacaaga agctctggaa gcagcagttg tagcaatgag
      421 agtgcttctg gttcaacttc cggtgaagca tccaacggtt gtcacaggtt cttgctctct
      481 cattctttct catcgtcttc ttcatcatct tcttcttcgt tgggagtttt tcctcgtaga
      541 ccagtgagat cagtagctaa aaaccccaaa tctgctccgg tagttagtaa acccttaatt
      601 agaaagaaac caagctctct tgaggaggtt aagttgaaat ctactcttac tgagaaacct
      661 aatttgctga agtctcagag atgcaagaca aatcctgtat ctggaaagag accaacttgt
      721 aagattacca tgaaaccgga gaaagtttcg gttttgaaga aacaatcttc tgtttctagc
      781 tagaactagt ggatccccca tcacaacttt gtatagaaaa gttgaacgag aaacgtaaaa
      841 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
      901 taaaacacaa catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt
      961 tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccggcgag attttcagga
     1021 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
     1081 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
     1141 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     1201 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attgcgatcg
     1261 catttaaatg gccggccctt aattaaggcg cgccgtttaa ggatcttcac ctagatcctt
     1321 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1381 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1441 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1501 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1561 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1621 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1681 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1741 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1801 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1861 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1921 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1981 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     2041 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2101 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2161 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2221 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2281 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2341 gttccgcgca catttccccg aaaagtgcca cctgaaacgg gcgacggatg gtgatccccc
     2401 tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata
     2461 tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta
     2521 tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc
     2581 tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgat
     2641 cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
     2701 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcaa ctttattata
     2761 caaagttgtg atatcaagct agcttcgaca attattttgg actctcatgt tctcaagctg
     2821 aaatccaaaa gaaacatgat gttggttcag atttggaatc actgacaatg aagtttcagt
     2881 cagccggttt gtcaccaagg atccaggctt catcatggga gccgagccgt tcaagtttca
     2941 acttcgacta tttggccaca tcttctgatt ctattgatct ctcacagttt cagagaggtt
     3001 tagttgacag gagttcttgt catcctcatg taacactgga taaagtgtca aggacacatt
     3061 tgagagtaga gcaaacaaat agccacatgc ctgaaatgaa atcccaacag attacagata
     3121 ctgaatttga tattcaaaat cacaaagaaa gtgctgcttc tctgggaaaa ggaagagaat
     3181 tgcttccatg ctctgctgca gagtcaataa gcactgatgg aggtggtctg atttgttctg
     3241 aggattcaaa ttggatggca tgttacaaga attacccagc tttcttgtac aaagttggca
     3301 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     3361 aaatcattat ttgccatcca gctgcagctc tggcccgtgt ctcaaaatct ctgatgttac
     3421 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     3481 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     3541 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     3601 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     3661 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     3721 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     3781 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     3841 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     3901 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     3961 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     4021 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     4081 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     4141 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     4201 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     4261 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     4321 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     4381 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4441 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4501 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4561 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4621 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4681 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4741 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4801 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4861 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4921 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4981 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5041 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5101 ttcctgcgtt atcccctgat tctgtggata accgtattac cgctagccag gaagagtttg
     5161 tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt
     5221 ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc
     5281 tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     5341 ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg
     5401 c
//
