LOCUS       40924_28531        8396 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLN-YC-Nano50 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8396)
  AUTHORS   Pandey K, Ferreira PE, Ishikawa T, Nagai T, Kaneko O, Yahata K.
  TITLE     Ca2+ monitoring in Plasmodium falciparum through yellow 
            cameleon-Nano biosensor
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8396)
  AUTHORS   Yahata K, Kaneko O.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-AUG-2015) Contact:Kazuhide Yahata Institute of 
            Tropical Medicine, Nagasaki University, Department of Parasitology; 
            1-12-4 Sakamoto, Nagasaki, Nagasaki 852-8042, Japan
REFERENCE   3  (bases 1 to 8396)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8396)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-AUG-2015) Contact:Kazuhide Yahata Institute of Tropical 
            Medicine, Nagasaki University, Department of Parasitology; 1-12-4 
            Sakamoto, Nagasaki, Nagasaki 852-8042, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8396
                     /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             2232..2309
                     /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             2532..3050
                     /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(3066..3086)
                     /label=attB3
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     primer_bind     complement(3094..3110)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4333..4437
                     /label=AmpR promoter
     CDS             4438..5295
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5469..6057
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6345..6366
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6381..6411
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6419..6435
                     /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     6443..6459
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6477..6495
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             7180..7572
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(8303..8320)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8327..8343)
                     /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 aggggggctc caagaggcgc tggaagaaaa acttcattgc cgtcagcgct gccaaccggt
     2281 tcaagaagat ctccagctcc ggggcactgg agctcatgga cggcggcgtg cagctcgccg
     2341 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2401 acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2461 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggtg
     2521 gcagcggtgg catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     2581 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     2641 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     2701 cctggcccac cctcgtgacc accctgggct acggcctgca gtgcttcgcc cgctaccccg
     2761 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2821 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2881 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2941 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     3001 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag ctcgagcccg
     3061 ggtaacaact ttattataca tagttgataa ttcactggcc gtcgttttac aacgtctcga
     3121 gggatatggc agcttaatgt tcgtttttct tatttatata tttataccaa ttgattgtat
     3181 ttataactgt aaaaatgtgt atgttgtgtg catatttttt tttgtgcatg cacatgcatg
     3241 taaatagcta aaattatgaa cattttattt tttgttcaga aaaaaaaaac tttacacaca
     3301 taaaatggct agtatgaata gccatatttt atataaatta aatcctatga atttatgacc
     3361 atattaaaaa tttagatatt tatggaacat aatatgtttg aaacaataag acaaaattat
     3421 tattattatt attattttta ctgttataat tatgtgtctc cttcaatgat tcataaatag
     3481 ttggacttga tttttaaaat gtttataata tgattagcat agttaaataa aaaaagttga
     3541 aaaattaaaa aaaaacatat aaacacaaat gatggttttt ccttcaattt cgatatcata
     3601 tagcaatcgt attagcaggt aaatgagcaa ggttaataaa aaaatggtaa aaaaaaaaaa
     3661 ttggggaaaa ataaacaagg tgtctttatc tacactttgg ctttacaatg tataatttat
     3721 atgcatacaa aaaaattttg ataaaattgt aaaaaggtga ataaaataac attgtaacag
     3781 tagtagatag taaagggaag gtgttgctca aatagtgtcg aaacaaaact ggcataaaga
     3841 aaaattgata ttgagcagag gatatgcgca taatggtatt ttgtttgttt gtttgttttc
     3901 tcattttttg agacagctca attctttatg tccacaacat catcggactt ttcttcttca
     3961 gggtaggcgg ccctgcagta cgtagcggtc tgggaagctc gcctgatgcg gtattttctc
     4021 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     4081 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4141 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4201 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4261 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4321 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4381 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4441 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4501 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4561 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4621 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4681 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4741 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4801 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4861 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4921 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4981 gtagcaatgc caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5041 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5101 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5161 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5221 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5281 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5341 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5401 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5461 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5521 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5581 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5641 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5701 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5761 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5821 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5881 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5941 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6001 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atcgaaaaac
     6061 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6121 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6181 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6241 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6301 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     6361 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6421 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctattt
     6481 aggtgacact atagaatact caagcttata attattaata ggtacttttt tttttatata
     6541 tgggaatttc cttatagggc ccgcatgctt agctaattcg cttgtaagag gtactctcgt
     6601 ttatgcaaaa ctatttgata tagcatttta acaagtacac atatatatat gtaatatata
     6661 tactatatat atctattgca tgtgtactaa gcatgtgcat ggcatcccct ttttctcgtg
     6721 tttaaaacag tttgtatgat aaaatataaa ggatttgaaa aagagaaaaa aatatatgat
     6781 ctcatcctat atagcgccat aatttttatt tgggttgaat aaaattttct actaaattta
     6841 ggtgtaagta aaataatgga atatatataa gtacaataaa aaagtgcata aattaaaaaa
     6901 tttttataat aaatattttt tttaaaaaag tcaataataa tattaaatat atataacaca
     6961 ggattatata tgttcactac aattttttat attataatat aaattctttt caattttcat
     7021 tttattttac atacactttc cttttttgtc actatatttt aatattcaca tatttagttt
     7081 aaatactggc tatttctttc tacatttgct agtaacaatt gtgtagtgct tatatatata
     7141 cacacaccta aaacttacaa accatggtga taaatgcatg ccaagccttt gtctcaagaa
     7201 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
     7261 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
     7321 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
     7381 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
     7441 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg
     7501 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat
     7561 gtgtgggagg gctaaggtac cccattaaat ttatttaata atagattaaa aatattataa
     7621 aaataaaaac ataaacacag aaattacaaa aaaaatacat atgaattttt tttttgtaat
     7681 cttccttata aatatagaat aatgaatcat ataaaacata tcatttttca tttatttaca
     7741 tttaaaatta ttgtttcagt atctttaatt tattatgtat atataaaaat aacttacaat
     7801 tttattaata aacaatatat gtttattaat tcatgttttg taatttatgg gatagcgatt
     7861 ttttttactg tctgtatttt tcttttttaa ttatgtttta attgtatttt atttttatta
     7921 ttgttctttt tatagtatta ttttaaaaca aaatgttttt tctaagaact tataataata
     7981 ataaatataa attttaataa aaattatatt tatcttttac aatatgaaca taaagtacaa
     8041 cattaatata tagcttttaa tatttttatt cctaatcatg taaatcttaa atttttcttt
     8101 ttaaacatat gttaaatatt tatttctcat tatatataag gacatattta ttaaaccgca
     8161 gagaaatcta gaggtaccga gctcgaattc cgggtttgta ccgtacacca ctgagaccgc
     8221 ggtggttgac cagacaaacc agaattcggg tttgtaccgt acaccactga gaccgcggtg
     8281 gttgaccaga caaaccagaa ttctatagtg agtcgtatta caattcactg gccgtcgttt
     8341 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagca
//
