LOCUS       sequence-211349-        6096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the ubiquitin ligase parkin fused to mTurquoise.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6096)
  AUTHORS   Nguyen J, Tirla A, Rivera-Fuentes P
  TITLE     Disruption of mitochondrial redox homeostasis by enzymatic 
            activation of a trialkylphosphine probe.
  JOURNAL   Org Biomol Chem. 2021 Mar 28;19(12):2681-2687. doi: 
            10.1039/d0ob02259d. Epub 2021 Feb 26.
  PUBMED    33634293
REFERENCE   2  (bases 1 to 6096)
  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: 
            "10.1039/d0ob02259d"; journalName: "Org Biomol Chem"; date: 
            "2021-03-28- 28"; volume: "19"; issue: "12"; pages: "2681-2687"
COMMENT     Sequence Label: pmTurquoise-parkin
FEATURES             Location/Qualifiers
     source          1..6096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        150..453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        454..657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     608..628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      696..705
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(702..704,705..707,708..1418)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(747..768)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1008..1027)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1355..1376
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2973..3094
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3010..3029)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3064..3083
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3101..3556)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3238..3259)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3450..3469
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3583..3687
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3689..4046
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3689..3880
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3712..3732)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3897..4032
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4081..4875
                     /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             4081..4875
                     /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(4135..4154)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4745..4764
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5063..5082)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5107..5154
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5483..6071
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5972..5991
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
       61 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
      121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
      661 ttagtgaacc gtcagatccg ctagcgctac cggtcgccac catggtgagc aagggcgagg
      721 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      781 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      841 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgtcct
      901 ggggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      961 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1021 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1081 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactact
     1141 ttagcgacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     1201 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca
     1261 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca
     1321 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1381 ccgccgggat cactctcggc atggacgagc tgtacaagtc cggactcaga tctcgagctc
     1441 aagctttgat agtgtttgtc aggttcaact ccagccatgg tttcccagtg gaggtcgatt
     1501 ctgacaccag catcttccag ctcaaggagg tggttgctaa gcgacagggg gttccggctg
     1561 accagttgcg tgtgattttc gcagggaagg agctgaggaa tgactggact gtgcagaatt
     1621 gtgacctgga tcagcagagc attgttcaca ttgtgcagag accgtggaga aaaggtcaag
     1681 aaatgaatgc aactggaggc gacgacccca gaaacgcggc gggaggctgt gagcgggagc
     1741 cccagagctt gactcgggtg gacctcagca gctcagtcct cccaggagac tctgtggggc
     1801 tggctgtcat tctgcacact gacagcagga aggactcacc accagctgga agtccagcag
     1861 gtagatcaat ctacaacagc ttttatgtgt attgcaaagg cccctgtcaa agagtgcagc
     1921 cgggaaaact cagggtacag tgcagcacct gcaggcaggc aacgctcacc ttgacccagg
     1981 gtccatcttg ctgggatgat gttttaattc caaaccggat gagtggtgaa tgccaatccc
     2041 cacactgccc tgggactagt gcagaatttt tctttaaatg tggagcacac cccacctctg
     2101 acaaggaaac atcagtagct ttgcacctga tcgcaacaaa tagtcggaac atcacttgca
     2161 ttacgtgcac agacgtcagg agccccgtcc tggttttcca gtgcaactcc cgccacgtga
     2221 tttgcttaga ctgtttccac ttatactgtg tgacaagact caatgatcgg cagtttgtcc
     2281 acgaccctca gcttggctac tccctgcctt gtgtggctgg ctgtcccaac tccttgatta
     2341 aagagctcca tcacttcagg attctgggag aagagcagta caaccggtac cagcagtatg
     2401 gtgcagagga gtgtgtcctg cagatggggg gcgtgttatg cccccgccct ggctgtggag
     2461 cggggctgct gccggagcct gaccagagga aagtcacctg cgaagggggc aatggcctgg
     2521 gctgtgggtt tgccttctgc cgggaatgta aagaagcgta ccatgaaggg gagtgcagtg
     2581 ccgtatttga agcctcagga acaactactc aggcctacag agtcgatgaa agagccgccg
     2641 agcaggctcg ttgggaagca gcctccaaag aaaccatcaa gaaaaccacc aagccctgtc
     2701 cccgctgcca tgtaccagtg gaaaaaaatg gaggctgcat gcacatgaag tgtccgcagc
     2761 cccagtgcag gctcgagtgg tgctggaact gtggctgcga gtggaaccgc gtctgcatgg
     2821 gggaccactg gttcgacgtg taaggatcca ccggatctag ataactgatc ataatcagcc
     2881 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2941 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3001 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3061 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     3121 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3181 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3241 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3301 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3361 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3421 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3481 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3541 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3601 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3661 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3721 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3781 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3841 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3901 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3961 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4021 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     4081 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4141 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4201 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4261 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4321 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4381 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4441 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4501 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4561 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4621 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4681 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4741 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4801 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4861 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4921 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4981 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5041 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     5101 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     5161 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5221 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5281 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5341 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5401 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5461 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5521 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5581 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5641 agccgtagtt aggccaccac ttcaagaact ctgtatcacc gcctacatac ctcgctctgc
     5701 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5761 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5821 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5881 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5941 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6001 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6061 gcctatggaa aaacgccagc aacgcggcct ttttac
//
