LOCUS       sequence-271910-        7527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal GFP-tagged mouse Trak1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7527)
  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 7527)
  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-Trak1
FEATURES             Location/Qualifiers
     source          1..7527
                     /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    3951..4072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3988..4007)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4042..4061
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4079..4534)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4216..4237)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4428..4447
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4561..4665
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4667..5024
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4667..4858
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4690..4710)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4875..5010
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5059..5853
                     /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             5059..5853
                     /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(5113..5132)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5723..5742
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6041..6060)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6085..6132
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6461..7049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6950..6969
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7224..7527
                     /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 cttcgaattc tatggcattg gcaattcagc
     1021 tcaggcagcc ctccagggcc caacctctgc caggactcag ccacacactg gctgggacag
     1081 actcatgtga cgtgtgcaac agcaccaacc ttccagaagt tgagatcatt agcctgctgg
     1141 aggaacagct gccccattac aagctaagag cggacaccat ctatggttac gaccacgatg
     1201 actggctgca tacgcccctc atctccccag atgccaacat cgacctcaca actgagcaga
     1261 tcgaagagac gctgaaatac ttccttctat gtgccgaaag agttggccag atgactaaga
     1321 catacaatga cattgatgct gtcacgaggc ttcttgagga gaaagagcgg gatttggagc
     1381 tggctgcgag gatcggtcag tcattgttga agaagaacaa gaccctaact gagaggaatg
     1441 aactgctgga agagcaggtg gagcacatcc gggaggaggt gtctcagctc cgacatgagc
     1501 tgtccatgaa agacgagctg cttcaattct acaccagtgc cgctgaggag agcgagcctg
     1561 agtccgtctg ctcaaccccg ctgaagagga acgagtcctc ctcctccgtc cagaattact
     1621 tccacctgga ctctcttcag aagaagctca aggacctcga agaggagaac gttgtacttc
     1681 gatccgaggc ctgtcagctg aagacagaga ccatcaccta tgaggagaag gagcagcagc
     1741 tggtgaacga ctgcgtgaag gagctgaggg atgccaacgt ccagattgcg agcatctctg
     1801 aggagctggc taagaagacg gaagatgccg cccgccagca ggaggagatc acccacctgc
     1861 tgtcgcaaat cgtggactta cagaagaagg caaaatcttg cgcggtggaa aatgaagagc
     1921 ttgtccagca cctgggcgct gccaaggacg cccagcgtca actcacagcc gagctccgag
     1981 agctggagga taagtacgcg gagtgcatgg agatgcttca tgaggctcag gaggagttga
     2041 agaacctgcg gaacaagacg atgcccacgt cccggcgcta ccactctctg ggcctgttcc
     2101 ccatggactc gttggctgca gaaatcgagg gaactatgcg caaggagctg cagctggaag
     2161 agctggagtc tccagacatc acccaccaga agcgtgtctt cgagacggtg aggaacgtca
     2221 atcaagtggt caagcagaga tcactaaccc cttctcccat gaacatcccg ggctccaacc
     2281 agtcctcagc catgaactcc ctcctgtcca gctgcgtcag cacccctcgc tccagcttct
     2341 acggcagcga cgtcagcaac gtggtcctcg ataacaaaac caacagcatc ctcctggaga
     2401 ccgaggcagc cgacctgggg aacgaggacc acaataagaa gccgggcact ccaggcacgc
     2461 caggctccca tgacctggag acggcactta ggcggctatc cctacgccgg gagaactacc
     2521 tgtctgagcg gaggttcttc gaggaggagc aggagaggaa gctgcgagag ctggctgaga
     2581 agggggagct gcacagtggc tcgctcacgc ccaccgagag catcatgtcc ctgggaacac
     2641 actcacgctt ctccgagttc acgggcttct ctggcatgtc cttcagcagc cgctcctacc
     2701 tgcccgagaa gctgcagatc gtgaagccac tcgaaggttc agccacgctt caccactggc
     2761 agcaattggc ccagcctcat cttgggggca tcctggaccc ccgtcctggt gtggtcacca
     2821 agggcttccg gactttggat gttgacctgg atgaagtgta ctgccttaac gactttgagg
     2881 aagatgacac aggtgaccac atttctctgg cgggcctagc tacctccacg ccaattcagc
     2941 acccggagac ctcagcgcac caccctggga agtgcatgtc acagaccaac tcaactttca
     3001 cctttaccac ttgccgcatc ctgcaccctt cagacgagct cactcgggtc acgccaagcc
     3061 ttaactctgc cccagctccg gcttgtagca gtaccagcca cttgaaatcc acaccagtgg
     3121 ccacgccatg caccccccgg agactgagcc tggctgagtc tttcacaaat gtccgtgagt
     3181 ccacgacaac catgagcacg tccctggggc tggtatggtt gctgaaggag cgaggcatct
     3241 cggcggccgt gtacgacccc cagagttggg acagagccgg caggggctcc ctcctccact
     3301 cctacacccc taggatggct gtgatcccgt ccaccccacc aaactcccct atgcagacac
     3361 cttcggcctc cccgccctcc ttcgagttca agtgcacgag ccctccctac aacaacttcc
     3421 tggcttccaa gccggccagc tctatcctga gggaggtgag ggaaaagagg cccgtgcgga
     3481 gcagcgagag ccaaacagat gtgtctgtct ccaacctcaa ccttgtggac aaagtcagga
     3541 ggtttggggt ggccagagtg gtgaactcag gacgtgcccg tatccccacc ttgactgagg
     3601 aacagggacc tctcctctgt ggacccacag ggccagcaca ggctcttgtc cctgggggcc
     3661 tggtccccga gggcctgcct ctggggtgcc ccagtggcat ccgacggaat cgcagcttcc
     3721 ccactatggt cgggtccagc gtgcagatga gagcccccgt gatcctcacc tcgggcatct
     3781 tgatgggtgc taagctcccc aaacagacta gcttgcggtg aggatccacc ggatctagat
     3841 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3901 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3961 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4021 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     4081 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4141 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4201 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4261 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4321 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4381 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4441 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4501 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     4561 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4621 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     4681 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4741 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4801 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4861 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4921 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4981 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     5041 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5101 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5161 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5221 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     5281 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5341 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5401 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5461 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5521 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5581 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5641 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5701 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5761 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5821 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5881 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5941 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     6001 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     6061 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     6121 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     6181 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     6241 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     6301 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6361 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6421 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6481 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6541 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6601 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6661 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6721 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6781 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6841 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6901 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6961 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7021 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7081 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7141 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     7201 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     7261 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     7321 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     7381 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     7441 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     7501 tacgtattag tcatcgctat taccatg
//
