LOCUS       Exported                9403 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Regulation of protein localization 
            (near-infrared-blue-light-inducible shuttle).
ACCESSION   .
VERSION     .
KEYWORDS    pQP-iRIS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9403)
  AUTHORS   Redchuk TA, Omelina ES, Chernov KG, Verkhusha VV
  TITLE     Near-infrared optogenetic pair for protein regulation and spectral 
            multiplexing.
  JOURNAL   Nat Chem Biol. 2017 Mar 27. doi: 10.1038/nchembio.2343.
  PUBMED    28346403
REFERENCE   2  (bases 1 to 9403)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2017 Mar 27. doi: 10.1038/nchembio.2343."
FEATURES             Location/Qualifiers
     source          1..9403
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      616..625
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2859..3569
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKL
                     ICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLER
                     VTAAGITLGMDELYK"
     primer_bind     complement(2898..2919)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3159..3178)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3624..3680
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    3682..4256
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3706..4258
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3850..3867)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4077..4096
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4315..5022
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4460..4478)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4728..4747
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5527..5541
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     polyA_signal    6191..6312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6228..6247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6282..6301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6319..6774)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6456..6477)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6668..6687
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6801..6905
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6907..7264
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(6930..6950)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7115..7250
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7177..7196
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7299..8093
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7353..7372)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7963..7982
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8281..8300)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8325..8372
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8701..9289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9190..9209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"