LOCUS       Exported                4741 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  photo-activatable mitochondrial marker for expression in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    mito-dendra2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4741)
  AUTHORS   Pham AH, McCaffery JM, Chan DC
  TITLE     Mouse lines with photo-activatable mitochondria to study 
            mitochondrial dynamics.
  JOURNAL   Genesis. 2012 Nov;50(11):833-43. doi: 10.1002/dvg.22050. Epub 2012 
            Aug 11.
  PUBMED    22821887
REFERENCE   2  (bases 1 to 4741)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1002/dvg.22050";
            journalName: "Genesis"; date: "2012-11"; volume: "50"; issue: "11"; 
            pages: "833-43"
FEATURES             Location/Qualifiers
     source          1..4741
                     /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"
     CDS             597..671
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             693..1379
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="NTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTAN
                     LTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKGI
                     CTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNINM
                     ALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARYS
                     PLPSQVW"
     polyA_signal    1529..1650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1566..1585)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1620..1639
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1657..2112)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1794..1815)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2006..2025
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2139..2243
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2245..2602
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2268..2288)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2453..2588
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2515..2534
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2637..3431
                     /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(2691..2710)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3301..3320
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3619..3638)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3663..3710
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4039..4627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4528..4547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"