LOCUS       sequence-384090-        6924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Mfn1 tagged with mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6924)
  AUTHORS   Nguyen TT, Voeltz GK
  TITLE     An ER phospholipid hydrolase drives ER-associated mitochondrial 
            constriction for fission and fusion.
  JOURNAL   Elife. 2022 Nov 30;11:e84279. doi: 10.7554/eLife.84279.
  PUBMED    36448541
REFERENCE   2  (bases 1 to 6924)
  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: "Elife."; 
            date: "2022-11-30"; volume: "11:e84279. doi"; pages: " 
            10.7554/eLife.84279"
COMMENT     Sequence Label: mNG-Mfn1
FEATURES             Location/Qualifiers
     source          1..6924
                     /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"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1320
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      868..877
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3712..3833
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3749..3768)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3803..3822
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3840..4295)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3977..3998)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4189..4208
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4322..4426
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4428..4785
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4428..4619
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4451..4471)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4636..4771
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4820..5614
                     /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             4820..5614
                     /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(4874..4893)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5484..5503
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5802..5821)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5846..5893
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6222..6810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6711..6730
                     /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 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggcctctct cccagcgaca
      661 catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt gggtcagggc
      721 accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa gggtgacctc
      781 cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca gtacctgccc
      841 taccctgacg ggatgtcgcc tttccaggcc gccatggtag atggctccgg ctaccaagtc
      901 catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg ctacacctac
      961 gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc tgctgacggt
     1021 cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa gacttacccc
     1081 aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa tggcaagcgc
     1141 taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc taactatctg
     1201 aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa gaccgagctc
     1261 aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga gctgtacaag
     1321 tccggactca gatctcgagc tcaagcttcg aattcgatgg cagaacctgt ttctccactg
     1381 aagcactttg tgctggctaa gaaggcgatt actgcaatct ttgaccagtt actggagttt
     1441 gttactgaag gatcacattt tgttgaagca acatataaga atccggaact tgatcgaata
     1501 gccactgaag atgatctggt agaaatgcaa ggatataaag acaagctttc catcattggt
     1561 gaggtgctat ctcggagaca catgaaggtg gcattttttg gcaggacaag cagtgggaag
     1621 agctctgtta tcaatgcaat gttgtgggat aaagttctcc ctagtgggat tggccatata
     1681 accaattgct tcctaagtgt tgaaggaact gatggagata aagcctatct tatgacagaa
     1741 ggatcagatg aaaaaaagag tgtgaagaca gttaatcaac tggcccatgc ccttcacatg
     1801 gacaaagatt tgaaagctgg ctgtcttgta cgtgtgtttt ggccaaaagc aaaatgtgcc
     1861 ctcttgagag atgacctggt gttagtagac agtccaggca cagatgtcac tacagagctg
     1921 gatagctgga ttgataagtt ttgcctagat gctgatgtct ttgttttggt cgcaaactct
     1981 gaatcaacac taatgaatac ggaaaaacac ttttttcaca aggtgaatga gcggctttcc
     2041 aagcctaata ttttcattct caataatcgt tgggatgcct ctgcatcaga gccagaatat
     2101 atggaagacg tacgcagaca gcacatggaa agatgcctgc atttcttggt ggaggagctc
     2161 aaagttgtaa atgctttaga agcacagaat cgtatcttct ttgtttcagc aaaggaagtt
     2221 cttagtgcta gaaagcaaaa agcacagggg atgccagaaa gtggtgtggc acttgctgaa
     2281 ggatttcatg caagattaca ggaatttcag aattttgaac aaatctttga ggagtgtatc
     2341 tcgcagtcag cagtgaaaac aaagttcgaa cagcacacta tcagagctaa acagatacta
     2401 gctactgtga aaaacataat ggattcagta aacctggcag ctgaagataa aaggcattat
     2461 tcagtggaag agagggaaga ccaaattgat agactggact ttattcgaaa ccagatgaac
     2521 cttttaacac tggatgttaa gaaaaaaatc aaggaggtta ccgaggaggt ggcaaacaaa
     2581 gtttcatgtg caatgacaga tgaaatttgt cgactgtctg ttttggttga tgaattttgt
     2641 tcagagtttc atcctaatcc agatgtatta aaaatatata aaagtgaatt aaataagcac
     2701 atagaggatg gtatgggaag aaatttggct gatcgatgca ccgatgaagt aaacgcctta
     2761 gtgcttcaga cccagcaaga aattattgaa aatttgaagc cattacttcc agctggtata
     2821 caggataaac tacatacact gatcccttgc aagaaatttg atctcagtta taatctaaat
     2881 taccacaagt tatgttcaga ttttcaagag gatattgtat ttcgtttttc cctgggctgg
     2941 tcttcccttg tacatcgatt tttgggccct agaaatgctc aaagggtgct cctaggatta
     3001 tcagagccta tctttcagct ccctagatct ttagcttcta ctcccactgc tcctaccact
     3061 ccagcaacgc cagataatgc atcacaggaa gaactcatga ttacattagt aacaggattg
     3121 gcgtccgtta catctagaac ttctatgggc atcattattg ttggaggagt gatttggaaa
     3181 actataggct ggaaactcct atctgtttca ttaactatgt atggagcttt gtatctttat
     3241 gaaagactga gctggaccac ccatgccaag gagcgagcct ttaaacagca gtttgtaaac
     3301 tatgcaactg aaaaactgag gatgattgtt agctccacga gtgcaaactg cagtcaccaa
     3361 gtaaaacaac aaatagctac cacttttgct cgcctgtgcc aacaagttga tattactcaa
     3421 aaacagctgg aagaagaaat tgctagatta cccaaagaaa tagatcagtt ggagaaaata
     3481 caaaacaatt caaagctctt aagaaataaa gctgttcaac ttgaaaatga gctggagaat
     3541 tttactaagc agtttctacc ttcaagcaat gaagaatcct aaggatccac cggatctaga
     3601 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3661 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3721 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3781 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     3841 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3901 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3961 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     4021 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     4081 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     4141 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     4201 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     4261 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     4321 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4381 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     4441 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4501 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4561 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4621 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4681 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4741 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     4801 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4861 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4921 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4981 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5041 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5101 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5161 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5221 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5281 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5341 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5401 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5461 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5521 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5581 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5641 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5701 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5761 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     5821 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5881 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5941 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     6001 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     6061 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6121 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6181 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6241 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6301 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6361 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6421 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6481 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6541 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6601 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6661 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6721 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6781 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6841 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6901 gataaccgta ttaccgccat gcat
//
