LOCUS       Exported                8396 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLN-YC-Nano50 DNA, complete sequence.
ACCESSION   LC075581
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8396
                     /organism="Expression vector pLN-YC-Nano50"
                     /mol_type="other DNA"
                     /db_xref="taxon:1707392"
     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..3065
                     /codon_start=1
                     /gene="YCNano50"
                     /product="yellow cameleon Nano50"
                     /label=YCNano50
                     /protein_id="BAU51804.1"
                     /translation="MYPAFLYKVAVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEG
                     DATYGKLTLKFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQ
                     ERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYIT
                     ADKQKNGIKAHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPN
                     EKRDHMVLLEFVTAARMHDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQN
                     PTEAELQDMINEVDADGNGTIYFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYIS
                     AAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAKGGSKRRWKKNFI
                     AVSAANRFKKISSSGALELMDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDP
                     NEKRDHMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHK
                     FSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFK
                     SAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYN
                     YNSHNVYITADKQKNGIKANFKIRHNIELEPG"
     gene            1062..3065
                     /gene="YCNano50"
                     /label=YCNano50
     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             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)
                     /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(3094..3110)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4333..4437
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4438..5298
                     /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      5469..6057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6345..6366
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /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     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             7174..7575
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MHAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFT
                     GVNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPG
                     IKAIVKDSDGQPTAVGIRELLPSGYVWEG"
     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"