LOCUS       Exported                4960 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Monomeric near-infrared fluorescent protein miRFP670 in mammalian 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    pmiRFP670-N1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4960)
  AUTHORS   Shcherbakova DM, Baloban M, Emelyanov AV, Brenowitz M, Guo P, 
            Verkhusha VV
  TITLE     Bright monomeric near-infrared fluorescent proteins as tags and 
            biosensors for multiscale imaging.
  JOURNAL   Nat Commun. 2016 Aug 19;7:12405. doi: 10.1038/ncomms12405.
  PUBMED    27539380
REFERENCE   2  (bases 1 to 4960)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2016-08-19"; volume: "7:12405. doi"; pages: " 
            10.1038/ncomms12405"
FEATURES             Location/Qualifiers
     source          1..4960
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        106..409
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        410..613
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     564..584
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    636..716
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      718..727
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             724..1671
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     polyA_signal    1793..1914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1830..1849)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1884..1903
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1921..2376)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2058..2079)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2270..2289
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2403..2507
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2509..2866
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2532..2552)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2717..2852
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2779..2798
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2901..3695
                     /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(2955..2974)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3565..3584
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3883..3902)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3927..3974
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4303..4891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4792..4811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"