LOCUS       40924_36568        8573 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway recycling vector pRED56-MD8-P35S DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8573)
  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 8573)
  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 8573)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8573)
  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..8573
                     /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        1582..1927
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    1949..2073
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2110..2140
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2194..2850
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             3195..3497
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3541..3665)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3697..3949
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    4158..4282
                     /label=attR4
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        4307..4337
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_feature    4609..4649
                     /label=rare cutter sites
                     /note="rare cutter sites"
     CDS             complement(4741..5598)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5599..5703)
                     /label=AmpR promoter
     protein_bind    complement(6000..6123)
                     /label=attR3
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(6449..6544)
                     /label=attL6
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             6668..7474
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      7620..8208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(8343..8370)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(8462..8548)
                     /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 aagcttgcat
     1081 gcctgcaggt ccccagatta gccttttcaa tttcagaaag aatgctaacc cacagatggt
     1141 tagagaggct tacgcagcag gtctcatcaa gacgatctac ccgagcaata atctccagga
     1201 aatcaaatac cttcccaaga aggttaaaga tgcagtcaaa agattcagga ctaactgcat
     1261 caagaacaca gagaaagata tatttctcaa gatcagaagt actattccag tatggacgat
     1321 tcaaggcttg cttcacaaac caaggcaagt aatagagatt ggagtctcta aaaaggtagt
     1381 tcccactgaa tcaaaggcca tggagtcaaa gattcaaata gaggacctaa cagaactcgc
     1441 cgtaaagact ggcgaacagt tcatacagag tctcttacga ctcaatgaca agaagaaaat
     1501 cttcgtcaac atggtggagc acgacacact tgtctactcc aaaaatatca aagatacagt
     1561 ctcagaagac caaagggcaa ttgagacttt tcaacaaagg gtaatatccg gaaacctcct
     1621 cggattccat tgcccagcta tctgtcactt tattgtgaag atagtggaaa aggaaggtgg
     1681 ctcctacaaa tgccatcatt gcgataaagg aaaggccatc gttgaagatg cctctgccga
     1741 cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc
     1801 aaccacgtct tcaaagcaag tggattgatg tgatatctcc actgacgtaa gggatgacgc
     1861 acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga
     1921 gagaacacgg gggactctag agttatcaac aagtttgtac aaaaaagctg aacgagaaac
     1981 gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac agactacata
     2041 atactgtaaa acacaacata tccagtcact atggcggccc ctcgaggggc cgcattaggc
     2101 accccaggct ttacacttta tgcttccggc tcgtataatg tgtggatttt gagttaggat
     2161 ccggcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc
     2221 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct
     2281 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag
     2341 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct
     2401 catccggaat tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac
     2461 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac
     2521 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa
     2581 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc
     2641 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc
     2701 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt
     2761 caggttcatc atgccgtctg tgatggcttc catgtcggca gaatgcttaa tgaattacaa
     2821 cagtactgcg atgagtggca gggcggggcg taaagatctg gatccggctt actaaaagcc
     2881 agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg
     2941 tatacccgaa gtatgtcaaa aagaggtgtg ctatgaagca gcgtattaca gtgacagttg
     3001 acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag
     3061 cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca
     3121 ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa
     3181 cagggactgg tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc
     3241 tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc
     3301 tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata
     3361 tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta
     3421 tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc
     3481 tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac
     3541 catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
     3601 taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa
     3661 gtggttgata acagcgctta gagctcgaat ttccccgatc gttcaaacat ttggcaataa
     3721 agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg
     3781 aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt
     3841 tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc
     3901 gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg ggaattctag
     3961 cttcgacaat tattttggac tctcatgttc tcaagctgaa atccaaaaga aacatgatgt
     4021 tggttcagat ttggaatcac tgacaatgaa gtttcagtca gccggtttgt caccaaggat
     4081 ccaggcttca tcatgggagc cgagccgttc aagtttcaac ttcgactatt tggatccgtc
     4141 ctagtggatc ccccatcaca actttgtata gaaaagttga acgagaaacg taaaatgata
     4201 taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa
     4261 cacaacatat ccagtcacta tggcggccgc attaggcacc ccaggcttta cactttatgc
     4321 ttccggctcg tataatgtgt ggattttgag ttaggatccg gcgagatttt caggagctaa
     4381 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     4441 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     4501 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     4561 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattgc gatcgcattt
     4621 aaatggccgg cccttaatta aggcgcgccg tttaaggatc ttcacctaga tccttttaaa
     4681 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4741 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4801 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4861 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4921 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4981 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5041 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5101 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5161 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5221 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5281 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5341 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5401 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5461 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5521 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5581 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5641 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5701 gcgcacattt ccccgaaaag tgccacctga aacgggcgac ggatggtgat ccccctggcc
     5761 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
     5821 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
     5881 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     5941 ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg tcgatcgacc
     6001 atagtgactg gatatgttgt gttttacagt attatgtagt ctgtttttta tgcaaaatct
     6061 aatttaatat attgatattt atatcatttt acgtttctcg ttcaacttta ttatacaaag
     6121 ttgtgatcca catcttctga ttctattgat ctctcacagt ttcagagagg tttagttgac
     6181 aggagttctt gtcatcctca tgtaacactg gataaagtgt caaggacaca tttgagagta
     6241 gagcaaacaa atagccacat gcctgaaatg aaatcccaac agattacaga tactgaattt
     6301 gatattcaaa atcacaaaga aagtgctgct tctctgggaa aaggaagaga attgcttcca
     6361 tgctctgctg cagagtcaat aagcactgat ggaggtggtc tgatttgttc tgaggattca
     6421 aattggatgg catgttacaa gaattaccca actttttaat acaaagttgg cattataaga
     6481 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     6541 atttgccatc cagctgcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     6601 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     6661 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     6721 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     6781 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     6841 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     6901 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     6961 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     7021 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     7081 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     7141 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     7201 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     7261 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     7321 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     7381 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     7441 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     7501 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     7561 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     7621 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     7681 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     7741 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     7801 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     7861 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7921 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7981 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     8041 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     8101 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     8161 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     8221 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     8281 ttatcccctg attctgtgga taaccgtatt accgctagcc aggaagagtt tgtagaaacg
     8341 caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg
     8401 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
     8461 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     8521 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgc
//
