LOCUS       sequence-271911-        6574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phospho-mutant of S156.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6574)
  AUTHORS   Birsa N, Norkett R, Wauer T, Mevissen TE, Wu HC, Foltynie T, Bhatia 
            K, Hirst WD, Komander D, Plun-Favreau H, Kittler JT
  TITLE     Lysine 27 ubiquitination of the mitochondrial transport protein Miro
            is dependent on serine 65 of the Parkin ubiquitin ligase.
  JOURNAL   J Biol Chem. 2014 May 23;289(21):14569-82. doi: 
            10.1074/jbc.M114.563031. Epub 2014 Mar 26.
  PUBMED    24671417
REFERENCE   2  (bases 1 to 6574)
  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"; doi: 
            "10.1074/jbc.M114.563031"; journalName: "J Biol Chem"; date: 
            "2014-05-23- 23"; volume: "289"; issue: "21"; pages: "14569-82"
COMMENT     Sequence Label: GFP-Miro1 S156A
FEATURES             Location/Qualifiers
     source          1..6574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(249..251,252..254,255..965)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(249..251,252..254,255..965)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(294..315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(555..574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     902..923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2998..3119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3035..3054)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3089..3108
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3126..3581)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3263..3284)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3475..3494
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3608..3712
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3714..4071
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3714..3905
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3737..3757)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3922..4057
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4106..4900
                     /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             4106..4900
                     /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(4160..4179)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4770..4789
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5088..5107)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5132..5179
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5508..6096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5997..6016
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6271..6574
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      301 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      361 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      421 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      481 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      541 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      601 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      661 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      721 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      781 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      841 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
      901 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      961 acaagtccgg actcagatct atgaagaaag acgtgcggat cctgctggtg ggagaaccta
     1021 gagttgggaa gacatcactg attatgtctc tggtcagtga agaatttcca gaagaggttc
     1081 ctccccgggc agaagaaatc accattccag ctgatgtcac cccagagaga gttccaacac
     1141 acattgtaga ttactcagaa gcagaacaga gtgatgaaca acttcatcaa gaaatatctc
     1201 aggctaatgt catctgtata gtgtatgccg ttaacaacaa gcattctatt gataaggtaa
     1261 caagtcgatg gattcctctc ataaatgaaa gaacagacaa agacagcagg ctgcctttaa
     1321 tattggttgg gaacaaatct gatctggtgg aatatagtag tatggagacc atccttccta
     1381 ttatgaacca gtatacagaa atagaaacct gtgtggagtg ttcagcgaaa aacctgaaga
     1441 acatagcaga gctcttttat tacgcacaga aagctgttct tcatcctaca gggcccctgt
     1501 actgcccaga ggagaaggag atgaaaccag cttgtataaa agcccttact cgtatattta
     1561 aaatatctga tcaagataat gatggtactc tcaatgatgc tgaactcaac ttctttcaga
     1621 ggatttgttt caacactcca ttagctcctc aagctctgga ggatgtcaag aatgtagtca
     1681 gaaaacatat aagtgatggt gtggctgaca gtgggttgac cctgaaaggt tttctctttt
     1741 tacacacact ttttatccag agagggagac acgaaactac ttggactgtg cttcgacgat
     1801 ttggttatga tgatgacctg gatttgacac ctgaatattt gttccccctg ctgaaaatac
     1861 ctcctgattg cactactgaa ttaaatcatc atgcatattt atttctccaa agcacctttg
     1921 acaagcatga tttggataga gactgtgctt tgtcacctga tgagcttaaa gatttattta
     1981 aagttttccc ttacatacct tgggggccag atgtgaataa cacagtttgt accaatgaaa
     2041 gaggctggat aacctaccag ggattccttt cccagtggac gctcacgact tatttagatg
     2101 tacagcggtg cctggaatat ttgggctatc taggctattc aatattgact gagcaagagt
     2161 ctcaagcttc agctgttaca gtgacaagag ataaaaagat agacctgcag aaaaaacaaa
     2221 ctcaaagaaa tgtgttcaga tgtaatgtaa ttggagtgaa aaactgtggg aaaagtggag
     2281 ttcttcaggc tcttcttgga agaaacttaa tgaggcagaa gaaaattcgt gaagatcata
     2341 aatcctacta tgcgattaac actgtttatg tatatggaca agagaaatac ttgttgttgc
     2401 atgatatctc agaatcggaa tttctaactg aagctgaaat catttgtgat gttgtatgcc
     2461 tggtatatga tgtcagcaat cccaaatcct ttgaatactg tgccaggatt tttaagcaac
     2521 actttatgga cagcagaata ccttgcttaa tcgtagctgc aaagtcagac ctgcatgaag
     2581 ttaaacaaga atacagtatt tcacctactg atttctgcag gaaacacaaa atgcctccac
     2641 cacaagcctt cacttgcaat actgctgatg cccccagtaa ggatatcttt gttaaattga
     2701 caacaatggc catgtatccg cacgtgacac aagctgacct caagagctcc acgttttggc
     2761 ttcgagcaag ttttggtgct actgtttttg cagttttggg ctttgctatg tacaaagcat
     2821 tattgaaaca gcgatgagaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga
     2881 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2941 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3001 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3061 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3121 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3181 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3241 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3301 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3361 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3421 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3481 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3541 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3601 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3661 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3721 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3781 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3841 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3901 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3961 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4021 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4081 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4141 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4201 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4261 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4321 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4381 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4441 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4501 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4561 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4621 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4681 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4741 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4801 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4861 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4921 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4981 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5041 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5101 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5161 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5221 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5281 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5341 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5401 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5461 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5521 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5581 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5641 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5701 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5761 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5821 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5881 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5941 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6001 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6061 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6121 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6181 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     6241 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     6301 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     6361 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     6421 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     6481 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     6541 gtacatctac gtattagtca tcgctattac catg
//
