LOCUS       sequence-437545-        5646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Peroxisome localization of PMP34 fused to mApple.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5646)
  AUTHORS   Li X, Gamuyao R, Wu ML, Cho WJ, King SV, Petersen RA, Stabley DR, 
            Lindow C, Climer LK, Shirinifard A, Ferrara F, Throm RE, Robinson 
            CG, Zhou Y, Carisey AF, Tebo AG, Chang CL
  TITLE     A fluorogenic complementation tool kit for interrogating lipid 
            droplet-organelle interaction.
  JOURNAL   J Cell Biol. 2024 Sep 2;223(9):e202311126. doi: 
            10.1083/jcb.202311126. Epub 2024 Jul 1.
  PUBMED    38949658
REFERENCE   2  (bases 1 to 5646)
  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."; date: "2024-09-2"; volume: "223(9):e202311126. doi"; pages: 
            " 10.1083/jcb.202311126. Epub 2024 Jul 1"
COMMENT     Sequence Label: PMP34-mApple
FEATURES             Location/Qualifiers
     source          1..5646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(49..70)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     261..280
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        394..498
                     /gene="bla"
                     /label=AmpR promoter
     promoter        500..857
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        500..691
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(523..543)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      708..843
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             892..1686
                     /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             892..1686
                     /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(946..965)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1556..1575
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1874..1893)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1918..1965
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2294..2882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2783..2802
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3057..3360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3361..3564
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3515..3535
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3591..3600
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4527..4536
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4533..5240
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     4946..4965
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5241..5306
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    5430..5551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5467..5486)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5521..5540
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(5558..5646,1..367))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
       61 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
      121 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
      181 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
      241 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
      301 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
      361 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
      421 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
      481 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
      541 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
      601 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
      661 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
      721 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
      781 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
      841 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
      901 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
      961 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     1021 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     1081 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     1141 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     1201 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     1261 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     1321 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     1381 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     1441 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     1501 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     1561 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     1621 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     1681 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     1741 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     1801 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     1861 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     1921 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     1981 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     2041 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     2101 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     2161 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2221 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2281 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2341 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2401 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2461 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2521 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2581 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2641 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2701 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2761 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2821 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2881 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2941 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     3001 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     3061 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     3121 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     3181 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     3241 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     3301 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     3361 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     3421 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     3481 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     3541 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gccaccatgg
     3601 cttccgtgct gtcctacgaa agcctggtcc acgccgtggc cggagccgtg ggaagcgtga
     3661 cagcaatgac agtgtttttt cccctggata cagctagact tcgacttcag gttgatgaga
     3721 aaagaaaatc caaaactaca cacatggtgc tcctggagat cattaaagaa gaaggactcc
     3781 tggcaccata tcgagggtgg tttccagtga tttccagtct ctgctgctcc aattttgtct
     3841 atttctacac ttttaatagc ctcaaagcac tctgggtcaa aggtcaacat tctaccactg
     3901 gaaaagatct ggtagttggg tttgttgcag gagtggttaa tgtgttgcta acaactccac
     3961 tctgggtggt aaacaccaga ctgaagcttc aaggagcaaa atttaggaat gaagacattg
     4021 taccaacaaa ctacaaaggt atcattgatg cttttcatca gatcattcgc gatgaaggaa
     4081 tctcggcttt atggaatggc acatttccct cattgctgtt ggtcttcaat cctgccatcc
     4141 agttcatgtt ttatgaaggt ttaaaacggc agcttttaaa gaaacggatg aagctttctt
     4201 ccttggatgt gttcatcatt ggtgcagtag ccaaagcgat tgccaccacg gtgacctatc
     4261 ccctgcagac ggtacagtca attctgaggt ttgggcgtca tagactaaac ccagaaaaca
     4321 gaacattggg aagtcttcgg aatattctct atcttcttca ccaacgagta agacgttttg
     4381 gaataatggg actctacaaa ggccttgaag ccaaactgct gcagacagtc ctcactgctg
     4441 ctctcatgtt ccttgtttat gagaaactga cagctgccac cttcacagtt atggggctga
     4501 agcgtgcaca ccaacaccta ccggtcgcca ccatggtgag caagggcgag gagaataaca
     4561 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     4621 acgagttcga gatcgagggc gagggcgagg gccgccccta cgaggccttt cagaccgcta
     4681 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     4741 tcatgtacgg ctccaaggtc tacattaagc acccagccga catccccgac tacttcaagc
     4801 tgtccttccc cgagggcttc aggtgggagc gcgtgatgaa cttcgaggac ggcggcatta
     4861 ttcacgttaa ccaggactcc tccctgcagg acggcgtgtt catctacaag gtgaagctgc
     4921 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     4981 cctccgagga gcggatgtac cccgaggacg gcgccctgaa gagcgagatc aagaagaggc
     5041 tgaagctgaa ggacggcggc cactacgccg ccgaggtcaa gaccacctac aaggccaaga
     5101 agcccgtgca gctgcccggc gcctacatcg tcgacatcaa gttggacatc gtgtcccaca
     5161 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     5221 gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcg aattctgcag
     5281 tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata
     5341 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     5401 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     5461 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5521 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     5581 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     5641 ggcaaa
//
