LOCUS       sequence-437546-        5956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Peroxisome localization of low-affinity NFAST fused to PMP34-Apple.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5956)
  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 5956)
  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-NFAST(low)
FEATURES             Location/Qualifiers
     source          1..5956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(430..1018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(510..529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1347..1394
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1419..1438
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1626..2420)
                     /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             complement(1626..2420)
                     /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(1737..1756)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2347..2366
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2455..2812)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2469..2604
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2621..2812
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2769..2789
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2814..2918)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2945..3400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3032..3051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3242..3263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3407..3528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3418..3437)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3472..3491
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4028..4735)
                     /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     complement(4303..4322)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      4732..4741
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5668..5677
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5704..5907)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5733..5753)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(5908..5956,1..255)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
       61 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
      121 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
      181 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc
      241 gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat
      301 tactattaat aactaatgca tggcggtaat acggttatcc acagaatcag gggataacgc
      361 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      421 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      481 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      541 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      601 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      661 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      721 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      781 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      841 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      901 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      961 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1021 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1081 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1141 aagttttaaa tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc
     1201 cccgacgttg gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa
     1261 ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca
     1321 gccctgggac cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg
     1381 tctttttatt gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt
     1441 tagcctcccc ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat
     1501 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga
     1561 atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt
     1621 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     1681 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     1741 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     1801 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     1861 gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct
     1921 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     1981 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     2041 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     2101 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     2161 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     2221 agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     2281 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     2341 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     2401 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca
     2461 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     2521 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     2581 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     2641 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     2701 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     2761 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc
     2821 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2881 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2941 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3001 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3061 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3121 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3181 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3241 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3301 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3361 tttaacgcga attttaacaa aatattaacg cttacaattt acgcgttaag atacattgat
     3421 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3481 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3541 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     3601 aacctctaca aatgtggtat ggctgattat gatctagagt cgcggccgct taggtcatgg
     3661 ccttcttcat gtgcaccttc accttggtgg ggcctctgct ggtggggatc atccactcga
     3721 acatggtgtt caggtcgccc ttctgcacgc cctccttgaa tctgccctgg aactccttgc
     3781 tctgggtgcc gggggccacg tcggtgaaga agttcttgcc gatcacggtc ttggggtctc
     3841 tgccggtgat ggtgccctcg gcggcgttgt agtggatgat cttgccgttg ccgtccagct
     3901 ggatggcgcc gaaggccagc ttgtccaggg ccttgtcgtc catcttggcc aggctgttct
     3961 cgatgtcgtc gccgccgaat ctcacggtct ccatggatcc ccctccgccg ctgccgcctc
     4021 ctccggactt gtacagctcg tccatgccgc cggtggagtg gcggccctcg gcgcgttcgt
     4081 actgttccac gatggtgtag tcctcgttgt gggacacgat gtccaacttg atgtcgacga
     4141 tgtaggcgcc gggcagctgc acgggcttct tggccttgta ggtggtcttg acctcggcgg
     4201 cgtagtggcc gccgtccttc agcttcagcc tcttcttgat ctcgctcttc agggcgccgt
     4261 cctcggggta catccgctcc tcggaggcct cccagcccat ggtcttcttc tgcattacgg
     4321 ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt gtagatgaac acgccgtcct
     4381 gcagggagga gtcctggtta acgtgaataa tgccgccgtc ctcgaagttc atcacgcgct
     4441 cccacctgaa gccctcgggg aaggacagct tgaagtagtc ggggatgtcg gctgggtgct
     4501 taatgtagac cttggagccg tacatgaact gaggggacag gatgtcccag gcgaagggca
     4561 gggggccacc cttggtcacc ttcagcttag cggtctgaaa ggcctcgtag gggcggccct
     4621 cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga gccctccatg tgcaccttga
     4681 agcgcatgaa ctccttgatg atggccatgt tattctcctc gcccttgctc accatggtgg
     4741 cgaccggtag gtgttggtgt gcacgcttca gccccataac tgtgaaggtg gcagctgtca
     4801 gtttctcata aacaaggaac atgagagcag cagtgaggac tgtctgcagc agtttggctt
     4861 caaggccttt gtagagtccc attattccaa aacgtcttac tcgttggtga agaagataga
     4921 gaatattccg aagacttccc aatgttctgt tttctgggtt tagtctatga cgcccaaacc
     4981 tcagaattga ctgtaccgtc tgcaggggat aggtcaccgt ggtggcaatc gctttggcta
     5041 ctgcaccaat gatgaacaca tccaaggaag aaagcttcat ccgtttcttt aaaagctgcc
     5101 gttttaaacc ttcataaaac atgaactgga tggcaggatt gaagaccaac agcaatgagg
     5161 gaaatgtgcc attccataaa gccgagattc cttcatcgcg aatgatctga tgaaaagcat
     5221 caatgatacc tttgtagttt gttggtacaa tgtcttcatt cctaaatttt gctccttgaa
     5281 gcttcagtct ggtgtttacc acccagagtg gagttgttag caacacatta accactcctg
     5341 caacaaaccc aactaccaga tcttttccag tggtagaatg ttgacctttg acccagagtg
     5401 ctttgaggct attaaaagtg tagaaataga caaaattgga gcagcagaga ctggaaatca
     5461 ctggaaacca ccctcgatat ggtgccagga gtccttcttc tttaatgatc tccaggagca
     5521 ccatgtgtgt agttttggat tttcttttct catcaacctg aagtcgaagt ctagctgtat
     5581 ccaggggaaa aaacactgtc attgctgtca cgcttcccac ggctccggcc acggcgtgga
     5641 ccaggctttc gtaggacagc acggaagcca tggtggctag cggatctgac ggttcactaa
     5701 accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg
     5761 ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc
     5821 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     5881 attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta
     5941 ctgccaagta ggaaag
//
