LOCUS       sequence-413173-        4887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of NbGFP-miRFP670nano3 localized to 
            mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4887)
  AUTHORS   Sekhon H, Ha JH, Presti MF, Procopio SB, Jarvis AR, Mirsky PO, John 
            AM, Loh SN
  TITLE     Adaptable, turn-on maturation (ATOM) fluorescent biosensors for 
            multiplexed detection in cells.
  JOURNAL   Nat Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w.
  PUBMED    37945909
REFERENCE   2  (bases 1 to 4887)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w."
COMMENT     Sequence Label: pCMV-Cox4-NbGFP_miRFP670nano3
FEATURES             Location/Qualifiers
     source          1..4887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     polyA_signal    1675..1796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1712..1731)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1766..1785
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1803..2258)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1940..1961)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2152..2171
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2285..2389
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2391..2748
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2391..2582
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2414..2434)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2599..2734
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2783..3577
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2783..3577
                     /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(2837..2856)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3447..3466
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3765..3784)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3809..3856
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4185..4773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4674..4693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgccac catgttgagt ctcaggcaat
      661 ccataaggtt cttcaagcct gcaacacgaa ctctttgttc tagtcggtat ctgctcgcac
      721 cggtctcagg tgctcaagtc caactggtgg agtctggtgg cgctttggtg cagccaggtg
      781 gctctctgcg tttgtcctgt gccgcttctg gcttcccagt gaaccgctat tccatgcgct
      841 ggtatcgcca ggctccaggc aaagagcgtg agtgggtagc cggtatgtcc agcgcgggtg
      901 atcgtagctc ctatgaagac tccggcggag ggggtagcat ggcaaacctg gacaagatgc
      961 tgaacaccac cgtgaccgag gtgcgcaagt tcctgcaagc ggacagagtg tgcgtgttca
     1021 agttcgagga agattactcc ggcaccgtct cgcacgaagc cgtggacgac agatggatta
     1081 gcatcctgaa gacccaggtg caggacagat acttcatgga aaccagaggc gaggaatacg
     1141 tccacggcag ataccaggcc atcgccgaca tctacacagc caatctggtc gagtgctaca
     1201 gagacctgct gatcgagttt caggtgcggg ccattctggc tgtccccatc ctgcaaggca
     1261 agaagctgtg gggcctgctg gtggcccacc aactggccgg ccctcgggag tggcagacct
     1321 gggaaatcga cttcctgaaa cagcaagccg tggtgatggg catcgccatc cagcagagcg
     1381 gcggaggggg tagcgtgaag ggccgtttca ccatcagccg tgacgatgcc cgtaacacgg
     1441 tgtatctgca aatgaacagc ttgaaacctg aagatacggc cgtgtattac tgtaatgtga
     1501 acgtgggctt cgagtattgg ggccaaggca cccaggtcac cgtctccagc taggcggccg
     1561 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     1621 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     1681 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     1741 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     1801 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     1861 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     1921 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     1981 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2041 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2101 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2161 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2221 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2281 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2341 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     2401 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2461 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2521 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2581 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2641 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     2701 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     2761 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     2821 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     2881 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     2941 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3001 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3061 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3121 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3181 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3241 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3301 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3361 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3421 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3481 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3541 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     3601 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3661 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     3721 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     3781 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     3841 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     3901 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     3961 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4021 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4081 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4141 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4201 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4261 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4321 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4381 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4441 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4501 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4561 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4621 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4681 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4741 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4801 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4861 gtggataacc gtattaccgc catgcat
//
