LOCUS       Exported                4539 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  N-peptide-mCherry expression vector pSA776, complete sequence.
ACCESSION   KM403568
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4539)
  AUTHORS   Auslander S, Stucheli P, Rehm C, Auslander D, Hartig JS, Fussenegger
            M.
  TITLE     A general design strategy for protein-responsive riboswitches in 
            mammalian cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4539)
  AUTHORS   Auslander S, Stucheli P, Rehm C, Auslander D, Hartig JS, Fussenegger
            M.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-AUG-2014) Department of Biosystems Science and 
            Engineering, D-BSSE, ETH Zurich, Mattenstrasse 26, Basel, BS 4058, 
            Switzerland
REFERENCE   3  (bases 1 to 4539)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4539
                     /organism="N-peptide-mCherry expression vector pSA776"
                     /mol_type="other DNA"
                     /db_xref="taxon:1549502"
     regulatory      37..245
                     /regulatory_class="promoter"
                     /gene="SV40 early promoter"
     gene            37..245
                     /gene="SV40 early promoter"
                     /label=SV40 early promoter
     promoter        48..244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      95..230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        286..304
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      322..331
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             328..1221
                     /codon_start=1
                     /gene="N-peptide-mCherry"
                     /product="N-peptide-mCherry"
                     /label=N-peptide-mCherry
                     /note="N-terminal fusion of N-peptide to red fluorescent 
                     protein mCherry"
                     /protein_id="AIR09511.1"
                     /translation="MDAQTRRRERRAEKQAQWKAANPLLVGVSAKPVNRPGGGSGGGGS
                     KLVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGP
                     LPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSS
                     LQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGG
                     HYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELY
                     KSGILQISSTVAAARV"
     gene            328..1221
                     /gene="N-peptide-mCherry"
                     /label=N-peptide-mCherry
                     /note="N-terminal fusion of N-peptide to red fluorescent 
                     protein mCherry"
     CDS             469..1173
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     polyA_signal    1256..1377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      1366..1371
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late mRNA polyadenylation signal"
     rep_origin      complement(1572..2215)
                     /direction=LEFT
                     /label=pUC ori
                     /note="pUC ori"
     rep_origin      complement(2042..2630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2801..3661)
                     /codon_start=1
                     /gene="AmpR"
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="AIR09512.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2801..3661)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(3662..3766)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3793..4248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    4379..4427
                     /note="synthetic polyadenylation signal"
     misc_feature    4441..4532
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"