LOCUS       Exported                6417 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  U1Ap-mCherry expression vector pSA906, complete sequence.
ACCESSION   KM403569
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6417)
  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  JOURNAL   Unpublished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 6417)
  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 6417)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6417
                     /organism="U1Ap-mCherry expression vector pSA906"
                     /mol_type="other DNA"
                     /db_xref="taxon:1549505"
     regulatory      232..819
                     /regulatory_class="promoter"
                     /gene="CMV promoter"
     gene            232..819
                     /gene="CMV promoter"
                     /label=CMV promoter
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      899..908
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             905..1984
                     /codon_start=1
                     /gene="U1Ap-mCherry"
                     /product="U1Ap-mCherry"
                     /label=U1Ap-mCherry
                     /note="N-terminal fusion protein of U1Ap (aa1-aa98) to the 
                     red fluorescent protein mCherry"
                     /protein_id="AIR09513.1"
                     /translation="MAVPETRPNHTIYINNLNEKIKKDELKKSLYAIFSQFGQILDILV
                     SRSLKMRGQAFVIFKEVSSATNALRSMQGFPFYDKPMRIQYAKTDSDIIAKMKGGGSGG
                     GGSKLVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTK
                     GGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQ
                     DSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLK
                     DGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMD
                     ELYKSGILQISSTVAAARV"
     gene            905..1984
                     /gene="U1Ap-mCherry"
                     /label=U1Ap-mCherry
                     /note="N-terminal fusion protein of U1Ap (aa1-aa98) to the 
                     red fluorescent protein mCherry"
     CDS             1232..1936
                     /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"
     regulatory      2017..2241
                     /regulatory_class="polyA_signal_sequence"
                     /note="BGH polyA"
     polyA_signal    2017..2241
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2287..2715
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     regulatory      2720..3063
                     /regulatory_class="promoter"
                     /gene="SV40 promoter"
     gene            2720..3063
                     /gene="SV40 promoter"
                     /label=SV40 promoter
     promoter        2729..3058
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2909..3044
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3125..3919
                     /codon_start=1
                     /gene="NeoR"
                     /product="NeoR"
                     /label=NeoR
                     /protein_id="AIR09514.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            3125..3919
                     /gene="NeoR"
                     /label=NeoR
     regulatory      4093..4223
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 polyA"
     polyA_signal    4093..4214
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4263..4279)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4287..4303
                     /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)."
     promoter        complement(4311..4341)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4356..4377
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      4606..5276
                     /label=pUC ori
                     /note="pUC ori"
     rep_origin      complement(4665..5250)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5421..6281)
                     /codon_start=1
                     /gene="AmpR"
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="AIR09515.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5421..6281)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(6282..6386)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(6323..6351)
                     /regulatory_class="promoter"
                     /gene="Pbla"
     gene            complement(6323..6351)
                     /gene="Pbla"
                     /label=Pbla