LOCUS       40924_36573        8017 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway recycling vector pRED56-MD8-Pnos DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8017)
  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 8017)
  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 8017)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8017)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Published 
            Only in Database (2012)"
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..8017
                     /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    670..1070
                     /label=MD8
                     /note="MD8"
     promoter        1191..1374
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     protein_bind    1393..1517
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1554..1584
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1638..2294
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2639..2941
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2985..3109)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3141..3393
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    3602..3726
                     /label=attR4
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3751..3781
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_feature    4053..4093
                     /label=rare cutter sites
                     /note="rare cutter sites"
     CDS             complement(4185..5042)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5043..5147)
                     /label=AmpR promoter
     protein_bind    complement(5444..5567)
                     /label=attR3
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(5893..5988)
                     /label=attL6
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             6112..6918
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      7064..7652
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7787..7814)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(7906..7992)
                     /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 ttcatcacac gtaatatcat ccattttata ttatgatcca caaacagcaa
      721 aggcccatta atctcattat attttagatt aattgattgc aattgcccaa cttctccact
      781 cagcaatgtt agttttaaag acacatcaag ttagtgtatt ttatttttaa tctttctata
      841 aaaaacaaca ctttgttttt gtctatttct tattctgtga aaaagagaaa tgaaaaagta
      901 aaaaagataa caaaaaggct ggtttttcac ggtgagaatt tttcatggtc acgcaataac
      961 ttaaatatga actattaaaa taactaagat tgaaaaatta gcatgatcaa aagaagaaga
     1021 aaaaaaagac aacatgaagc ggaaaggaaa aaaaagaaag ctgaaaatct aagcttatca
     1081 tgagcggaga attaagggag tcacgttatg acccccgccg atgacgcggg acaagccgtt
     1141 ttacgtttgg aactgacaga accgcaacgt tgaaggagcc actcagccgc gggtttctgg
     1201 agtttaatga gctaagcaca tacgtcagaa accattattg cgcgttcaaa agtcgcctaa
     1261 ggtcactatc agctagcaaa tatttcttgt caaaaatgct ccactgacgt tccataaatt
     1321 cccctcggta tccaattaga gtctcatatt cactctcaat ccaaataatc tgcaccggat
     1381 ctagagttat caacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat
     1441 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     1501 catatccagt cactatggcg gcccctcgag gggccgcatt aggcacccca ggctttacac
     1561 tttatgcttc cggctcgtat aatgtgtgga ttttgagtta ggatccggcg agattttcag
     1621 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     1681 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     1741 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     1801 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta
     1861 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     1921 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     1981 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     2041 ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca
     2101 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     2161 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     2221 tctgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     2281 ggcagggcgg ggcgtaaaga tctggatccg gcttactaaa agccagataa cagtatgcgt
     2341 atttgcgcgc tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt
     2401 caaaaagagg tgtgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca
     2461 gttgctcaag gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat
     2521 gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag
     2581 gtcgcccggt ttattgaaat gaacggctct tttgctgacg agaacaggga ctggtgaaat
     2641 gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca
     2701 gagtgatatt attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct
     2761 gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg
     2821 gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc
     2881 tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat
     2941 ataaatgtca ggctccctta tacacagcca gtctgcaggt cgaccatagt gactggatat
     3001 gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga
     3061 tatttatatc attttacgtt tctcgttcag ctttcttgta caaagtggtt gataacagcg
     3121 cttagagctc gaatttcccc gatcgttcaa acatttggca ataaagtttc ttaagattga
     3181 atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg
     3241 taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc
     3301 cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat
     3361 tatcgcgcgc ggtgtcatct atgttactag atcgggaatt ctagcttcga caattatttt
     3421 ggactctcat gttctcaagc tgaaatccaa aagaaacatg atgttggttc agatttggaa
     3481 tcactgacaa tgaagtttca gtcagccggt ttgtcaccaa ggatccaggc ttcatcatgg
     3541 gagccgagcc gttcaagttt caacttcgac tatttggatc cgtcctagtg gatcccccat
     3601 cacaactttg tatagaaaag ttgaacgaga aacgtaaaat gatataaata tcaatatatt
     3661 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
     3721 actatggcgg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
     3781 gtgtggattt tgagttagga tccggcgaga ttttcaggag ctaaggaagc taaaatggag
     3841 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     3901 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     3961 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     4021 cttgcccgcc tgatgaatgc tcatccggaa ttgcgatcgc atttaaatgg ccggccctta
     4081 attaaggcgc gccgtttaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4141 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4201 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4261 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4321 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4381 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4441 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     4501 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4561 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4621 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4681 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4741 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     4801 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4861 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4921 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4981 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5041 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5101 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5161 aaagtgccac ctgaaacggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt
     5221 cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
     5281 tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc
     5341 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa
     5401 tgtcaggctc ccttatacac agccagtctg caggtcgatc gaccatagtg actggatatg
     5461 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
     5521 atttatatca ttttacgttt ctcgttcaac tttattatac aaagttgtga tccacatctt
     5581 ctgattctat tgatctctca cagtttcaga gaggtttagt tgacaggagt tcttgtcatc
     5641 ctcatgtaac actggataaa gtgtcaagga cacatttgag agtagagcaa acaaatagcc
     5701 acatgcctga aatgaaatcc caacagatta cagatactga atttgatatt caaaatcaca
     5761 aagaaagtgc tgcttctctg ggaaaaggaa gagaattgct tccatgctct gctgcagagt
     5821 caataagcac tgatggaggt ggtctgattt gttctgagga ttcaaattgg atggcatgtt
     5881 acaagaatta cccaactttt taatacaaag ttggcattat aagaaagcat tgcttatcaa
     5941 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     6001 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     6061 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     6121 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     6181 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     6241 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     6301 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     6361 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     6421 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     6481 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     6541 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     6601 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     6661 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     6721 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     6781 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     6841 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     6901 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     6961 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttttcgt
     7021 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     7081 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7141 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7201 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7261 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7321 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7381 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7441 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7501 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7561 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7621 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7681 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7741 tggataaccg tattaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     7801 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     7861 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     7921 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     7981 tcgttttatt tgatgcctgg cagttcccta ctctcgc
//
