LOCUS       sequence-271914-        7474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal GFP-tagged mouse Trak2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7474)
  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 7474)
  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-Trak2
FEATURES             Location/Qualifiers
     source          1..7474
                     /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    3898..4019
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3935..3954)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3989..4008
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4026..4481)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4163..4184)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4375..4394
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4508..4612
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4614..4971
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4614..4805
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4637..4657)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4822..4957
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5006..5800
                     /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             5006..5800
                     /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(5060..5079)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5670..5689
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5988..6007)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6032..6079
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6408..6996
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6897..6916
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7171..7474
                     /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 cgagctcaag cttcgatgag tcaatcccag aatgccattt
     1021 tcaaatcacg aacaggtgaa gcaaacctca tgagtagcaa tcatagagac tcggagagca
     1081 tcactgatgt ctgctccaac gaggatctcc ctgaagtcga gctggtgaac ttgctagaag
     1141 aacagctgcc acagtataag ctaagagtag attctctttt tctctatgaa aaccaagact
     1201 gggcacagtc atcacaccag cagcaggatg cgcctgagac cctctcccca gtccttgctg
     1261 aagagacgtt ccgctacatg attctaggca cagacagagt ggagcagatg accaaaacct
     1321 acaatgacat tgacatggtc acacatctcc tggcagagag ggatcgagat ctagagctag
     1381 ctgcccgaat tgggcaagct cttttaaagc gaaaccatgt cttatctgag cagaatgaag
     1441 ctctggagga gcagttggga caagcctttg atcaagttaa tcagctgcag cacgagctct
     1501 ccaagaaaga cgagctgctc cgaatcgtct ctattgcctc cgaggagagt gaaacggatt
     1561 ctagctgctc cacgcccctt cggttcaatg agtccttcag cttatctcag gggctgctgc
     1621 agctggacat gctgcatgag aagctcaggg agctggaaga agagaacatg gccctacgat
     1681 ccaaggcttg tcatatcaag acagaaacat ttacctatga agagaaggaa caacagctcg
     1741 tcaatgactg tgttaaagaa cttcgtgaga ccaatgcaca gatgtccaga atgactgaag
     1801 agctgtcggg aaagagcgac gagctgctcc ggtaccaaga ggagatctcc tccctgctgt
     1861 ctcagatcgt agaccttcag cacaaactga aagagcatgt gatcgagaag gaagaactaa
     1921 gacttcacct gcaggcttcc aaagatgccc agcgacagct gacaatggag cttcatgagt
     1981 tacaggacag gaacatggaa tgcctgggaa tgttacatga gtcccaagaa gaaataaaag
     2041 agcttcgtag caagtccggc ccttctgctc atctctgctt ctcccagtct tacggcgttt
     2101 ttacagggga atcgttggca gctgagattg aagggaccat gcgtaaaaag ctgagtttgg
     2161 atgaggaatc tgtgtctaaa caaaaggccc agcagaaacg ggtgtttgat actgtcaagg
     2221 ttgccaatga cactcggggc cgctctgtca ccttcccagt cctgctaccc atcccaggct
     2281 ccaaccgttc aagtgtcatc atgacagcaa agccctttga gtctggtgtc cagccagcag
     2341 aggacaaaac actcctgagc ccagggggca gcacagaggt tccaggaaac tcgcagccca
     2401 caaacccccc aggatcccct gaagatagtg atctggctac tgcattgcat cgcctcagcc
     2461 tgagaagaca gaactaccta agtgagaagc agttctttgc tgaggaatgg gaacggaaga
     2521 tccagatctt ggctgagcag gaggaagaag tcagcagctg tgacgccccc acggagaacc
     2581 ttgcctcagt ctgcactgac cagtcggaga ccacggacct cggcagtgcc agctgccttc
     2641 gaggcttcat gccagagaag ttacagattg tcaagcccct agaaggatca cagactctgc
     2701 atcactggca gcagcttgct cagccgaact tgggaaccat tcttgatcca cgcccaggcg
     2761 tcatcactaa aggcttcaca caggtgccca aggatgttgt ctaccacatc tcggacttag
     2821 aggaggatga agaagagggc atcacttttc aggttcagca gcctcttcaa ctggagcaga
     2881 aacctgcact gcccacgcca gtaacgggaa tctttctgcc gcccatgacc tcagcgggtg
     2941 gaccaggtac agttgcaact tcgaacccag gaaagtgtct ctcattcacc aactcgacat
     3001 tcaccttcac cacctgcagg atcttacatc cctctgacat cactcaggtc acccctagct
     3061 ccggtttccc atcactggcc tgtggcagta gtgcgagcag ctcatccaac acggctgtga
     3121 actcccctgc tgcatcctat agactaagca ttggcgaatc catcacaaac cgcagggact
     3181 ccaccataac cttcagtagc accaggagct tagccaaact cctgcaggag cgaggcatct
     3241 ctgctaaagt gtaccacagc ccggcttcag aaaaccccct cctgcagcct tgccccaagg
     3301 ccctggccac cccctccaca ccaccaaatt ccccagcgca gtcaccgtgc tcctccccct
     3361 tgccctttga accccgagtg catgtctccg agaatttctt ggcttcccga ccagctgaaa
     3421 cattcctgca agaaatgtat ggcttgagac cttcaagggc ccctcctgat gttggacagc
     3481 tgaagatgaa cttggtagac aggctgaaga ggctgggaat agccagggtg gtcaagaccc
     3541 ctgatcccca ggagaacgga aaaagccgag aagcagaaat gggtcttcaa aaaccagacc
     3601 ctgctgttta tttaaattca ggtggcagtt tattgtgtgg actgaggagg aatcagagtc
     3661 ttccggtcat gatgggtagc tttggagccc cagtttgcac aacctcaccc aaaatgggta
     3721 tcctgaagga agattgagaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga
     3781 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3841 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3901 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3961 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     4021 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     4081 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     4141 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     4201 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     4261 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     4321 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     4381 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     4441 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     4501 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4561 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     4621 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4681 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4741 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4801 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4861 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4921 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4981 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     5041 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5101 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5161 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     5221 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5281 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5341 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5401 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5461 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5521 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5581 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5641 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5701 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5761 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5821 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5881 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5941 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     6001 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     6061 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     6121 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     6181 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     6241 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6301 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6361 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6421 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6481 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6541 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6601 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6661 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6721 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6781 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6841 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6901 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6961 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7021 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7081 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     7141 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     7201 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     7261 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     7321 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     7381 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     7441 gtacatctac gtattagtca tcgctattac catg
//
