LOCUS       40924_36558        7934 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway recycling vector pRED56-MD8-CY2 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7934)
  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  JOURNAL   Unpublished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 7934)
  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 7934)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7934)
  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            "Unpublished"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..7934
                     /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    1109..1233
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1270..1300
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1354..2010
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2355..2657
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2701..2825)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2836..3036
                     /codon_start=1
                     /gene="ceyfp"
                     /product="carboxy-terminal fragment of enhanced yellow
                     fluorescent protein"
                     /label=ceyfp
                     /protein_id="BAT33537.1"
                     /translation="MGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDH
                     MVLLEFVTAAGITLGMDELYK"
     gene            2836..3036
                     /gene="ceyfp"
                     /label=ceyfp
     terminator      3058..3310
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    3519..3643
                     /label=attR4
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3668..3698
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_feature    3970..4010
                     /label=rare cutter sites
                     /note="rare cutter sites"
     CDS             complement(4102..4959)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4960..5064)
                     /label=AmpR promoter
     protein_bind    complement(5361..5484)
                     /label=attR3
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(5810..5905)
                     /label=attL6
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             6029..6835
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      6981..7569
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7704..7731)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(7823..7909)
                     /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 taagcttgca
     1081 tgcctgcagg tcgactctag agttatcaac aagtttgtac aaaaaagctg aacgagaaac
     1141 gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac agactacata
     1201 atactgtaaa acacaacata tccagtcact atggcggccc ctcgaggggc cgcattaggc
     1261 accccaggct ttacacttta tgcttccggc tcgtataatg tgtggatttt gagttaggat
     1321 ccgtcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc
     1381 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct
     1441 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag
     1501 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct
     1561 catccggaat tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac
     1621 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac
     1681 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa
     1741 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc
     1801 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc
     1861 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt
     1921 caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa
     1981 cagtactgcg atgagtggca gggcggggcg taaagatctg gatccggctt actaaaagcc
     2041 agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg
     2101 tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca gtgacagttg
     2161 acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag
     2221 cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca
     2281 ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa
     2341 caggggctgg tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc
     2401 tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc
     2461 tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata
     2521 tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta
     2581 tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc
     2641 tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac
     2701 catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
     2761 taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa
     2821 gtggttgata acagcatggg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     2881 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg
     2941 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3001 gggatcactc tcggcatgga cgagctgtac aagtaagctt agagctcgaa tttccccgat
     3061 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg
     3121 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg
     3181 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg
     3241 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg
     3301 ttactagatc gggaattcta gcttcgacaa ttattttgga ctctcatgtt ctcaagctga
     3361 aatccaaaag aaacgtgatg ttggttcaga tttggaatca ctgacaatga agtttcagtc
     3421 agccggtttg tcaccaagga tccaggcttc atcatgggag ccgagccgtt caagtttcaa
     3481 cttcgactat ttggatccgt cctagtggat cccccatcac aactttgtat agaaaagttg
     3541 aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac
     3601 agactacata atactgtaaa acacaacata tccagtcact atggcggccg cattaggcac
     3661 cccaggcttt acactttatg cttccggctc gtataatgtg tggattttga gttaggatcc
     3721 ggcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac
     3781 cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca
     3841 atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa
     3901 aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca
     3961 tccggaattg cgatcgcatt taaatggccg gcccttaatt aaggcgcgcc gtttaaggat
     4021 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4081 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4141 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4201 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4261 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4321 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4381 agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc
     4441 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4501 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4561 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4621 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4681 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag
     4741 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4801 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4861 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4921 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4981 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5041 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aaacgggcga
     5101 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt
     5161 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
     5221 gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
     5281 aaaaacgcca ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc
     5341 cagtctgcag gtcgatcgac catagtgact ggatatgttg tgttttacag tattatgtag
     5401 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     5461 gttcaacttt attatacaaa gttgtgatcc acatcttctg attctattga tctctcacag
     5521 tttcagagag gtttagttga caggagttct tgtcatcctc atgtaacact ggataaagtg
     5581 tcaaggacac atttgagagt agagcaaaca aatagccaca tgcctgaaat gaaatcccaa
     5641 cagattacag atactgaatt tgatattcaa aatcacaaag aaagtgctgc ttctctggga
     5701 aaaggaagag aattgcttcc atgctctgct gcagagtcaa taagcactga tggaggtggt
     5761 ctgatttgtt ctgaggattc aaattggatg gcatgttaca agaattaccc aactttttaa
     5821 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     5881 atcagtcaaa ataaaatcat tatttgccat ccagctgcag ctctggcccg tgtctcaaaa
     5941 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     6001 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     6061 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     6121 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     6181 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     6241 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     6301 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     6361 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     6421 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     6481 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     6541 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     6601 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     6661 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     6721 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     6781 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     6841 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     6901 aagctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6961 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7021 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7081 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     7141 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7201 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7261 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7321 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7381 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7441 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7501 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7561 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7621 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgctagc
     7681 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
     7741 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
     7801 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     7861 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     7921 ttccctactc tcgc
//
