LOCUS       Exported                8402 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLN-YC Nano15 DNA, complete sequence.
ACCESSION   LC028929
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8402
                     /organism="Cloning vector pLN-YC Nano15"
                     /mol_type="other DNA"
                     /db_xref="taxon:1617784"
     protein_bind    152..172
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1029..1053
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1062..3071
                     /codon_start=1
                     /gene="YCNano15"
                     /product="yellow cameleon Nano15"
                     /label=YCNano15
                     /protein_id="BAU51803.1"
                     /translation="MYPAFLYKVAVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEG
                     DATYGKLTLKFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQ
                     ERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYIT
                     ADKQKNGIKAHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPN
                     EKRDHMVLLEFVTAARMHDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQN
                     PTEAELQDMINEVDADGNGTIYFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYIS
                     AAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAKGGGGSKRRWKKN
                     FIAVSAANRFKKISSSGALELMDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSK
                     DPNEKRDHMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNG
                     HKFSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDF
                     FKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLE
                     YNYNSHNVYITADKQKNGIKANFKIRHNIELEPG"
     gene            1062..3071
                     /gene="YCNano15"
                     /label=YCNano15
     protein_bind    complement(1066..1090)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     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)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /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
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4444..5304
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5475..6063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6351..6372
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /bound_moiety="lac repressor encoded by lacI"
                     /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             7180..7581
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MHAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFT
                     GVNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPG
                     IKAIVKDSDGQPTAVGIRELLPSGYVWEG"
     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"