LOCUS       sequence-404955-        7562 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid of GFP-tagged hOpa1 isoform 1 mutant 
            protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7562)
  AUTHORS   Nyenhuis SB, Wu X, Strub MP, Yim YI, Stanton AE, Baena V, Syed ZA, 
            Canagarajah B, Hammer JA, Hinshaw JE
  TITLE     OPA1 helical structures give perspective to mitochondrial 
            dysfunction.
  JOURNAL   Nature. 2023 Aug 23. doi: 10.1038/s41586-023-06462-1.
  PUBMED    37612506
REFERENCE   2  (bases 1 to 7562)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2023 Aug 23. doi: 10.1038/s41586-023-06462-1."
COMMENT     Sequence Label: Control-2
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     75..95
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3058..3067
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3064..3066,3067..3069,3070..3783)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3109..3130)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3370..3389)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3717..3738
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3906..4027
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3943..3962)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3997..4016
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4034..4489)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4171..4192)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4383..4402
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4516..4620
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4622..4979
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4622..4813
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4645..4665)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4830..4965
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5014..5808
                     /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             5014..5808
                     /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(5068..5087)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5678..5697
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5996..6015)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6040..6087
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6416..7004
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6905..6924
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7179..7482
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7483..7562,1..124)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
       61 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      121 agctggttta gtgaaccgtc agatccgcta gcgctaccga tgtggcgact acgtcgggcc
      181 gctgtggcct gtgaggtctg ccagtcttta gtgaaacaca gctctggaat aaaaggaagt
      241 ttaccactac aaaaactaca tctggtttca cgaagcattt atcattcaca tcatcctacc
      301 ttaaagcttc aacgacccca attaaggaca tcctttcagc agttctcttc tctgacaaac
      361 cttcctttac gtaaactgaa attctctcca attaaatatg gctaccagcc tcgcaggaat
      421 ttttggccag caagattagc tacgagactc ttaaaacttc gctatctcat actaggatcg
      481 gctgttgggg gtggctacac agccaaaaag acttttgatc agtggaaaga tatgataccg
      541 gaccttagtg aatataaatg gattgtgcct gacattgtgt gggaaattga tgagtatatc
      601 gattttgaga aaattagaaa agcccttcct aattcagaag accttgtaaa gttagcacca
      661 gactttgaca agattgttga aagccttagc ttattgaagg acttttttac ctcaggttct
      721 ccggaagaaa cggcgtttag agcaacagat cgtggatctg aaagtgacaa gcattttaga
      781 aaggtgtcag acaaagagaa aattgaccaa cttcaggaag aacttctgca cactcagttg
      841 aagtatcaga gaatcttgga acgattagaa aaggagaaca aagaattgag aaaattagta
      901 gcccagaaag atgacaaagg cattcatcat agaaagctta agaaatcttt gattgacatg
      961 tattctgaag ttcttgatgt tctctctgat tatgatgcca gttataatac gcaagatcat
     1021 ctgccacggg ttgttgtggt tggagatcag agtgctggaa agactagtgt gttggaaatg
     1081 attgcccaag ctcgaatatt cccaagagga tctggggaga tgatgacacg ttctccagtt
     1141 aaggtgactc tgagtgaagg tcctcaccat gtggccctat ttaaagatag ttctcgggag
     1201 tttgatctta ccaaagaaga agatcttgca gcattaagac atgaaataga acttcgaatg
     1261 aggaaaaatg tgaaagaagg ctgtaccgtt agccctgaga ccatatcctt aaatgtaaaa
     1321 ggccctggac tacagaggat ggtgcttgtt gacttaccag gtgtgattaa tactgtgaca
     1381 tcaggcatgg ctcctgacac aaaggaaact attttcagta tcagcaaagc ttacatgcag
     1441 aatcctaatg ccatcatact gtgtattcaa gatggatctg tggatgctga acgcagtatt
     1501 gttacagact tggtcagtca aatggaccct catggaagga gaaccatatt cgttttgacc
     1561 aaagtagacc tggcagagaa aaatgtagcc agtccaagca ggattcagca gataattgaa
     1621 ggaaagctct tcccaatgaa agctttaggt tattttgctg ttgtaacagg aaaagggaac
     1681 agctctgaaa gcattgaagc tataagagaa tatgaagaag agttttttca gaattcaaag
     1741 ctcctaaaga caagcatgct aaaggcacac caagtgacta caagaaattt aagccttgca
     1801 gtatcagact gcttttggaa aatggtacga gagtctgttg aacaacaggc tgatagtttc
     1861 aaagcaacac gttttaacct tgaaactgaa tggaagaata actatcctcg cctgcgggaa
     1921 cttgaccgga atgaactatt tgaaaaagct aaaaatgaaa tccttgatga agttatcagt
     1981 ctgagccagg ttacaccaaa acattgggag gaaatccttc aacaatcttt gtgggaaaga
     2041 gtatcaactc atgtgattga aaacatctac cttccagctg cgcagaccat gaattcagga
     2101 acttttaaca ccacagtgga tatcaagctt aaacagtgga ctgataaaca acttcctaat
     2161 aaagcagtag aggttgcttg ggagacccta caagaagaat tttcccgctt tatgacagaa
     2221 ccgaaaggga aagagcatga tgacatattt gataaactta aagaggccgt taaggaagaa
     2281 agtattaaac gacacaagtg gaatgacttt gcggaggaca gcttgagggt tattcaacac
     2341 aatgctttgg aagaccgatc catatctgat aaacagcaat gggatgcagc tatttatttt
     2401 atggaagagg ctctgcaggc tcgtctcaag gatactgaaa atgcaattga aaacatggtg
     2461 ggtccagact ggaaaaagag gtggttatac tggaagaatc ggacccaaga acagtgtgtt
     2521 cacaatgaaa ccaagaatga attggagaag atgttgaaat gtaatgagga gcacccagct
     2581 tatcttgcaa gtgatgaaat aaccacagtc cggaagaacc ttgaatcccg aggagtagaa
     2641 gtagatccaa gcttgattaa ggatacttgg catcaagttt atagaagaca ttttttaaaa
     2701 acagctctaa accattgtaa cctttgtcga agaggttttt attactacca aaggcatttt
     2761 gtagattctg agttggaatg caatgatgtg gtcttgtttt ggcgtataca gcgcatgctt
     2821 gctatcaccg caaatacttt aaggcaacaa cttacaaata ctgaagttag gcgattagag
     2881 aaaaatgtta aagaggtatt ggaagatttt gctgaagatg gtgagaagaa gattaaattg
     2941 cttactggta aacgcgttca actggcggaa gacctcaaga aagttagaga aattcaagaa
     3001 aaacttgatg ctttcattga agctcttcat caggagaaag ggctagcgct accggtcgcc
     3061 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     3121 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     3181 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     3241 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     3301 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     3361 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     3421 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3481 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     3541 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     3601 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3661 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     3721 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3781 taaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     3841 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3901 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3961 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4021 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     4081 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4141 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4201 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     4261 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     4321 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4381 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4441 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     4501 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4561 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4621 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4681 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4741 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4801 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4861 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4921 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     4981 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5041 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5101 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5161 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     5221 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5281 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5341 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5401 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5461 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5521 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     5581 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5641 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5701 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5761 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5821 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5881 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5941 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     6001 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     6061 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     6121 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     6181 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     6241 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     6301 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6361 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6421 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6481 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6541 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6601 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6661 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6721 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6781 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6841 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6901 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6961 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7021 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     7081 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
     7141 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     7201 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     7261 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     7321 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     7381 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     7441 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     7501 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     7561 ga
//
