LOCUS       sequence-418611-        6096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CFP tagged Parkin for colocalization study.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6096)
  AUTHORS   Narendra D, Tanaka A, Suen DF, Youle RJ
  TITLE     Parkin is recruited selectively to impaired mitochondria and 
            promotes their autophagy.
  JOURNAL   J Cell Biol. 2008 Dec 1. 183(5):795-803.
  PUBMED    19029340
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"; journalName: "J Cell 
            Biol. 2008 Dec 1. 183(5):795-803."
COMMENT     Sequence Label: CFP-Parkin
FEATURES             Location/Qualifiers
     source          1..6096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        142..445
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        446..649
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     600..620
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(694..696,697..699,700..1410)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(739..760)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1000..1019)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1347..1368
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2965..3086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3002..3021)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3056..3075
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3093..3548)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3230..3251)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3442..3461
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3575..3679
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3681..4038
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3681..3872
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3704..3724)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3889..4024
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4073..4867
                     /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             4073..4867
                     /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(4127..4146)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4737..4756
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5055..5074)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5099..5146
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5475..6063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5964..5983
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
       61 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
      121 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      181 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      241 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      301 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      361 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      421 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      481 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      541 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      601 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      661 ccgtcagatc cgctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc
      721 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      781 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
      841 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg
      901 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      961 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1021 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1081 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac
     1141 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     1201 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1261 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1321 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1381 atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc tcaagctttg
     1441 atagtgtttg tcaggttcaa ctccagccat ggtttcccag tggaggtcga ttctgacacc
     1501 agcatcttcc agctcaagga ggtggttgct aagcgacagg gggttccggc tgaccagttg
     1561 cgtgtgattt tcgcagggaa ggagctgagg aatgactgga ctgtgcagaa ttgtgacctg
     1621 gatcagcaga gcattgttca cattgtgcag agaccgtgga gaaaaggtca agaaatgaat
     1681 gcaactggag gcgacgaccc cagaaacgcg gcgggaggct gtgagcggga gccccagagc
     1741 ttgactcggg tggacctcag cagctcagtc ctcccaggag actctgtggg gctggctgtc
     1801 attctgcaca ctgacagcag gaaggactca ccaccagctg gaagtccagc aggtagatca
     1861 atctacaaca gcttttatgt gtattgcaaa ggcccctgtc aaagagtgca gccgggaaaa
     1921 ctcagggtac agtgcagcac ctgcaggcag gcaacgctca ccttgaccca gggtccatct
     1981 tgctgggatg atgttttaat tccaaaccgg atgagtggtg aatgccaatc cccacactgc
     2041 cctgggacta gtgcagaatt tttctttaaa tgtggagcac accccacctc tgacaaggaa
     2101 acatcagtag ctttgcacct gatcgcaaca aatagtcgga acatcacttg cattacgtgc
     2161 acagacgtca ggagccccgt cctggttttc cagtgcaact cccgccacgt gatttgctta
     2221 gactgtttcc acttatactg tgtgacaaga ctcaatgatc ggcagtttgt ccacgaccct
     2281 cagcttggct actccctgcc ttgtgtggct ggctgtccca actccttgat taaagagctc
     2341 catcacttca ggattctggg agaagagcag tacaaccggt accagcagta tggtgcagag
     2401 gagtgtgtcc tgcagatggg gggcgtgtta tgcccccgcc ctggctgtgg agcggggctg
     2461 ctgccggagc ctgaccagag gaaagtcacc tgcgaagggg gcaatggcct gggctgtggg
     2521 tttgccttct gccgggaatg taaagaagcg taccatgaag gggagtgcag tgccgtattt
     2581 gaagcctcag gaacaactac tcaggcctac agagtcgatg aaagagccgc cgagcaggct
     2641 cgttgggaag cagcctccaa agaaaccatc aagaaaacca ccaagccctg tccccgctgc
     2701 catgtaccag tggaaaaaaa tggaggctgc atgcacatga agtgtccgca gccccagtgc
     2761 aggctcgagt ggtgctggaa ctgtggctgc gagtggaacc gcgtctgcat gggggaccac
     2821 tggttcgacg tgtaaggatc caccggatct agataactga tcataatcag ccataccaca
     2881 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     2941 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3001 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3061 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     3121 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3181 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3241 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3301 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3361 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3421 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3481 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3541 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3601 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3661 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3721 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3781 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3841 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3901 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3961 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4021 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4081 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4141 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4201 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4261 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4321 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4381 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4441 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4501 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4561 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4621 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4681 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4741 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4801 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4861 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4921 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4981 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     5041 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5101 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5161 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5221 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5281 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5341 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5401 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5461 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5521 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5581 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5641 ttaggccacc acttcaagaa ctctgtatca ccgcctacat acctcgctct gctaatcctg
     5701 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5761 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5821 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5881 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5941 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6001 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6061 aaaaacgcca gcaacgcggc ctttttacgg ttcctg
//
