LOCUS       40924_28526        8402 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLN-YC Nano15 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8402)
  AUTHORS   Yahata K.
  TITLE     Yellow Cameleon-Nano biosensors for calcium monitoring against 
            inhibitors for sarco/endoplasmic reticulum Ca2+-ATPase in Plasmodium
            falciparum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8402)
  AUTHORS   Yahata K.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-FEB-2015) Contact:Kazuhide Yahata Institute of 
            Tropical Medicine, Protozoology; 1-12-4 Sakamoto, Nagasaki, Nagasaki
            852-8523, Japan URL 
            :http://www.tm.nagasaki-u.ac.jp/protozoology/index.html
REFERENCE   3  (bases 1 to 8402)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8402)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-FEB-2015) Contact:Kazuhide Yahata Institute of Tropical 
            Medicine, Protozoology"; volume: " 1-12-4 Sakamoto, Nagasaki, 
            Nagasaki 852-8523, Japan URL :http"; pages: 
            "//www.tm.nagasaki-u.ac.jp/protozoology/index.htm"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    152..172
                     /label=attB4
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     protein_bind    1029..1053
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1066..1090)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1092..1772
                     /codon_start=1
                     /label=GFP
                     /note="green fluorescent protein"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     HFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAA"
     CDS             2238..2315
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             2538..3056
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     protein_bind    complement(3072..3092)
                     /label=attB3
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     primer_bind     complement(3100..3116)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4339..4443
                     /label=AmpR promoter
     CDS             4444..5301
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5475..6063
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6351..6372
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6387..6417
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6425..6441
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6449..6465
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6483..6501
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             7186..7578
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(8309..8326)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8333..8349)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
       61 cagttgcgca gcctgaatgg cgaatggcga gcttgggaat actcaagctg cggcccgggt
      121 accagctatg accatgatta cgccaagcta tcaactttgt atagaaaagt tgtcgtcgac
      181 ggaaaggggc cattggatat atatttagta ttccatagta tatagttaaa tataataata
      241 ttatagtaaa tttttgcatg aatttaaaga aaaaaaaaaa aaaaataaca aaaaggagta
      301 ttatataaat ataacaaaat aataaatact atacctatat atatatatat atatatatac
      361 atatatataa atatataaat atatttatat aaaaataata ttttatatat aatattatat
      421 ttttataata tctttttttt ttttaaatat catctttctt tctttctttc tttttttttt
      481 tttgatttat atatwatttt attttgtatt tacaaaatat aataaaataa tttcatgttt
      541 agcaaaaatt ttaaataraa aataataata taaaataaaa agcatcattt aatataaaat
      601 aataaaatta aaataaaata agataattac caaaaataat aaaaaaaatt aaaaagtatt
      661 aaaaaatata tatcatatat aaatatataa attttaacaa cgtgagaaat taagaaaaaa
      721 attatattta tataaataat ataatattta atatatatct catataatat tatatatata
      781 tatatatata tatatttttt aaaaaaaaag aaaaaataag tttattttta ccttttaata
      841 aaaacatatt tttagatata cataaaaatt tttatattat tttattggta tatatatata
      901 tatatatata tatataataa tatatatata ataaaatata taaatattta cataaatatt
      961 ttactttaaa aagaaatttt ccttttttat tatatttttt ttcttcccac atttcgaata
     1021 aaagatctgc aagtttgtac aaaaaagcag gctattcaaa aatgtaccca gctttcttgt
     1081 acaaagtggc tgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1141 agctggacgg cgacgtaaac ggccacaggt tcagcgtgtc cggcgagggc gagggcgatg
     1201 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     1261 ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg cttcagccgc taccccgacc
     1321 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgta
     1381 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1441 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1501 tggggcacaa gctggagtac aactacatca gccacaacgt ctatatcacc gccgacaagc
     1561 agaagaacgg catcaaggcc cacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     1621 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1681 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     1741 acatggtcct gctggagttc gtgaccgccg cccgcatgca tgaccaactg acagaagagc
     1801 agattgcaga gttcaaagaa gccttctcat tattcgacaa ggatggggac ggcaccatca
     1861 ccacaaagga acttggcacc gttatgaggt cgcttggaca aaacccaacg gaagcagaat
     1921 tgcaggatat gatcaatgaa gtcgatgctg atggcaatgg aacgatttac tttcctgaat
     1981 ttcttactat gatggctaga aaaatgaagg acacagacag cgaagaggaa atccgagaag
     2041 cattccgtgt ttttgacaag gatgggaacg gctacatcag cgctgctgaa ttacgtcacg
     2101 tcatgacaaa cctcggggag aagttaacag atgaagaagt tgatgaaatg ataagggaag
     2161 cagatatcga tggtgatggc caagtaaact atgaagagtt tgtacaaatg atgacagcaa
     2221 aggggggagg tggctccaag aggcgctgga agaaaaactt cattgccgtc agcgctgcca
     2281 accggttcaa gaagatctcc agctccgggg cactggagct catggacggc ggcgtgcagc
     2341 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2401 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     2461 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2521 agggtggcag cggtggcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2581 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     2641 agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc
     2701 ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct
     2761 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2821 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     2881 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2941 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg
     3001 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgagctcg
     3061 agcccgggta acaactttat tatacatagt tgataattca ctggccgtcg ttttacaacg
     3121 tctcgaggga tatggcagct taatgttcgt ttttcttatt tatatattta taccaattga
     3181 ttgtatttat aactgtaaaa atgtgtatgt tgtgtgcata tttttttttg tgcatgcaca
     3241 tgcatgtaaa tagctaaaat tatgaacatt ttattttttg ttcagaaaaa aaaaacttta
     3301 cacacataaa atggctagta tgaatagcca tattttatat aaattaaatc ctatgaattt
     3361 atgaccatat taaaaattta gatatttatg gaacataata tgtttgaaac aataagacaa
     3421 aattattatt attattatta tttttactgt tataattatg tgtctccttc aatgattcat
     3481 aaatagttgg acttgatttt taaaatgttt ataatatgat tagcatagtt aaataaaaaa
     3541 agttgaaaaa ttaaaaaaaa acatataaac acaaatgatg gtttttcctt caatttcgat
     3601 atcatatagc aatcgtatta gcaggtaaat gagcaaggtt aataaaaaaa tggtaaaaaa
     3661 aaaaaattgg ggaaaaataa acaaggtgtc tttatctaca ctttggcttt acaatgtata
     3721 atttatatgc atacaaaaaa attttgataa aattgtaaaa aggtgaataa aataacattg
     3781 taacagtagt agatagtaaa gggaaggtgt tgctcaaata gtgtcgaaac aaaactggca
     3841 taaagaaaaa ttgatattga gcagaggata tgcgcataat ggtattttgt ttgtttgttt
     3901 gttttctcat tttttgagac agctcaattc tttatgtcca caacatcatc ggacttttct
     3961 tcttcagggt aggcggccct gcagtacgta gcggtctggg aagctcgcct gatgcggtat
     4021 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     4081 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     4141 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4201 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     4261 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4321 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4381 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4441 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4501 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4561 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4621 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4681 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4741 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4801 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4861 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4921 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4981 atgcctgtag caatgccaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5041 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5101 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5161 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5221 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5281 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5341 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5401 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5461 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5521 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5581 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     5641 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5701 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5761 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5821 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5881 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5941 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6001 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatcg
     6061 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6121 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6181 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6241 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6301 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     6361 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     6421 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     6481 ctatttaggt gacactatag aatactcaag cttataatta ttaataggta cttttttttt
     6541 tatatatggg aatttcctta tagggcccgc atgcttagct aattcgcttg taagaggtac
     6601 tctcgtttat gcaaaactat ttgatatagc attttaacaa gtacacatat atatatgtaa
     6661 tatatatact atatatatct attgcatgtg tactaagcat gtgcatggca tccccttttt
     6721 ctcgtgttta aaacagtttg tatgataaaa tataaaggat ttgaaaaaga gaaaaaaata
     6781 tatgatctca tcctatatag cgccataatt tttatttggg ttgaataaaa ttttctacta
     6841 aatttaggtg taagtaaaat aatggaatat atataagtac aataaaaaag tgcataaatt
     6901 aaaaaatttt tataataaat atttttttta aaaaagtcaa taataatatt aaatatatat
     6961 aacacaggat tatatatgtt cactacaatt ttttatatta taatataaat tcttttcaat
     7021 tttcatttta ttttacatac actttccttt tttgtcacta tattttaata ttcacatatt
     7081 tagtttaaat actggctatt tctttctaca tttgctagta acaattgtgt agtgcttata
     7141 tatatacaca cacctaaaac ttacaaacca tggtgataaa tgcatgccaa gcctttgtct
     7201 caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct
     7261 gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc
     7321 aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct
     7381 gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc
     7441 ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc
     7501 atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct
     7561 ggttatgtgt gggagggcta aggtacccca ttaaatttat ttaataatag attaaaaata
     7621 ttataaaaat aaaaacataa acacagaaat tacaaaaaaa atacatatga attttttttt
     7681 tgtaatcttc cttataaata tagaataatg aatcatataa aacatatcat ttttcattta
     7741 tttacattta aaattattgt ttcagtatct ttaatttatt atgtatatat aaaaataact
     7801 tacaatttta ttaataaaca atatatgttt attaattcat gttttgtaat ttatgggata
     7861 gcgatttttt ttactgtctg tatttttctt ttttaattat gttttaattg tattttattt
     7921 ttattattgt tctttttata gtattatttt aaaacaaaat gttttttcta agaacttata
     7981 ataataataa atataaattt taataaaaat tatatttatc ttttacaata tgaacataaa
     8041 gtacaacatt aatatatagc ttttaatatt tttattccta atcatgtaaa tcttaaattt
     8101 ttctttttaa acatatgtta aatatttatt tctcattata tataaggaca tatttattaa
     8161 accgcagaga aatctagagg taccgagctc gaattccggg tttgtaccgt acaccactga
     8221 gaccgcggtg gttgaccaga caaaccagaa ttcgggtttg taccgtacac cactgagacc
     8281 gcggtggttg accagacaaa ccagaattct atagtgagtc gtattacaat tcactggccg
     8341 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     8401 ca
//
