LOCUS       sequence-404939-        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: Interface-1 (e)
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2930..2939
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2936..2938,2939..2941,2942..3655)
                     /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(2981..3002)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3242..3261)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3589..3610
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3778..3899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3815..3834)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3869..3888
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3906..4361)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4043..4064)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4255..4274
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4388..4492
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4494..4851
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4494..4685
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4517..4537)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4702..4837
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4886..5680
                     /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             4886..5680
                     /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(4940..4959)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5550..5569
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5868..5887)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5912..5959
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6288..6876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6777..6796
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7051..7354
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7355..7558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7509..7529
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 tagtgaaccg tcagatccgc tagcgctacc gatgtggcga ctacgtcggg ccgctgtggc
       61 ctgtgaggtc tgccagtctt tagtgaaaca cagctctgga ataaaaggaa gtttaccact
      121 acaaaaacta catctggttt cacgaagcat ttatcattca catcatccta ccttaaagct
      181 tcaacgaccc caattaagga catcctttca gcagttctct tctctgacaa accttccttt
      241 acgtaaactg aaattctctc caattaaata tggctaccag cctcgcagga atttttggcc
      301 agcaagatta gctacgagac tcttaaaact tcgctatctc atactaggat cggctgttgg
      361 gggtggctac acagccaaaa agacttttga tcagtggaaa gatatgatac cggaccttag
      421 tgaatataaa tggattgtgc ctgacattgt gtgggaaatt gatgagtata tcgattttga
      481 gaaaattaga aaagcccttc ctaattcaga agaccttgta aagttagcac cagactttga
      541 caagattgtt gaaagcctta gcttattgaa ggactttttt acctcaggtt ctccggaaga
      601 aacggcgttt agagcaacag atcgtggatc tgaaagtgac aagcatttta gaaaggtgtc
      661 agacaaagag aaaattgacc aacttcagga agaacttctg cacactcagt tgaagtatca
      721 gagaatcttg gaacgattag aaaaggagaa caaagaattg agaaaattag tattgcagaa
      781 agatgacaaa ggcattcatc atagaaagct taagaaatct ttgattgaca tgtattctga
      841 agttcttgat gttctctctg attatgatgc cagttataat acgcaagatc atctgccacg
      901 ggttgttgtg gttggagatc agagtgctgg aaagactagt gtgttggaaa tgattgccca
      961 agctcgaata ttcccaagag gatctgggga gatgatgaca cgttctccag ttaaggtgac
     1021 tctgagtgaa ggtcctcacc atgtggccct atttaaagat agttctcggg agtttgatct
     1081 taccaaagaa gaagatcttg cagcattaag acatgaaata gaacttcgaa tgaggaaaaa
     1141 tgtgaaagaa ggctgtaccg ttagccctga gaccatatcc ttaaatgtaa aaggccctgg
     1201 actacagagg atggtgcttg ttgacttacc aggtgtgatt aatactgtga catcaggcat
     1261 ggctcctgac acaaaggaaa ctattttcag tatcagcaaa gcttacatgc agaatcctaa
     1321 tgccatcata ctgtgtattc aagatggatc tgtggatgct gaacgcagta ttgttacaga
     1381 cttggtcagt caaatggacc ctcatggaag gagaaccata ttcgttttga ccaaagtaga
     1441 cctggcagag aaaaatgtag ccagtccaag caggattcag cagataattg aaggaaagct
     1501 cttcccaatg aaagctttag gttattttgc tgttgtaaca ggaaaaggga acagctctga
     1561 aagcattgaa gctataagag aatatgaaga agagtttttt cagaattcaa agctcctaaa
     1621 gacaagcatg ctaaaggcac accaagtgac tacaagaaat ttaagccttg cagtatcaga
     1681 ctgcttttgg aaaatggtac gagagtctgt tgaacaacag gctgatagtt tcaaagcaac
     1741 acgttttaac cttgaaactg aatggaagaa taactatcct cgcctgcggg aacttgaccg
     1801 gaatgaacta tttgaaaaag ctaaaaatga aatccttgat gaagttatca gtctgagcca
     1861 ggttacacca aaacattggg aggaaatcct tcaacaatct ttgtgggaaa gagtatcaac
     1921 tgcggtgatt gaaaacatct accttccagc tgcgcagacc atgaattcag gaacttttaa
     1981 caccacagtg gatatcaagc ttaaacagtg gactgataaa caacttccta ataaagcagt
     2041 agaggttgct tgggagaccc tacaagaaga attttcccgc tttatgacag aaccgaaagg
     2101 gaaagagcat gatgacatat ttgataaact taaagaggcc gttaaggaag aaagtattaa
     2161 acgacacaag tggaatgact ttgcggagga cagcttgagg gttattcaac acaatgcttt
     2221 ggaagaccga tccatatctg ataaacagca atgggatgca gctatttatt ttatggaaga
     2281 ggctctgcag gctcgtctca aggatactga aaatgcaatt gaaaacatgg tgggtccaga
     2341 ctggaaaaag aggtggttat actggaagaa tcggacccaa gaacagtgtg ttcacaatga
     2401 aaccaagaat gaattggaga agatgttgaa atgtaatgag gagcacccag cttatcttgc
     2461 aagtgatgaa ataaccacag tccggaagaa ccttgaatcc cgaggagtag aagtagatcc
     2521 aagcttgatt aaggatactt ggcatcaagt ttatagaaga cattttttaa aaacagctct
     2581 aaaccattgt aacctttgtc gaagaggttt ttattactac caaaggcatt ttgtagattc
     2641 tgagttggaa tgcaatgatg tggtcttgtt ttggcgtata cagcgcatgc ttgctatcac
     2701 cgcaaatact ttaaggcaac aacttacaaa tactgaagtt aggcgattag agaaaaatgt
     2761 taaagaggta ttggaagatt ttgctgaaga tggtgagaag aagattaaat tgcttactgg
     2821 taaacgcgtt caactggcgg aagacctcaa gaaagttaga gaaattcaag aaaaacttga
     2881 tgctttcatt gaagctcttc atcaggagaa agggctagcg ctaccggtcg ccaccatggt
     2941 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     3001 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     3061 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     3121 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     3181 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3241 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3301 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3361 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     3421 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     3481 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3541 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3601 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg
     3661 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3721 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3781 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3841 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3901 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3961 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     4021 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     4081 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     4141 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     4201 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     4261 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     4321 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     4381 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4441 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     4501 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4561 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4621 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4681 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4741 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4801 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4861 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4921 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4981 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5041 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     5101 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5161 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5221 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5281 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5341 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5401 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5461 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5521 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5581 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5641 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5701 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5761 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5821 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5881 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5941 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     6001 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     6061 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     6121 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6181 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6241 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6301 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6361 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6421 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6481 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6541 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6601 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6661 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6721 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6781 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6841 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6901 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6961 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     7021 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     7081 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     7141 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     7201 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     7261 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     7321 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     7381 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     7441 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     7501 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     7561 tt
//
