LOCUS       sequence-426360-        8442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Detection of peroxisome-lipid droplet contact site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8442)
  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 8442)
  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: FAB(PX-LD)
FEATURES             Location/Qualifiers
     source          1..8442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(77..665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(157..176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(836..1627,1628..1696))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1459..1478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1697..1801)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1876..1895)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2067..2446
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2447..2650
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2601..2621
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2695..2714
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2695..2713
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2763..2772
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3699..3708
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3753..4073
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             4104..4145
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          4622..4851
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     4834..4853
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(4923..5497,5498..5500,5501..5509)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5090..5107)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5317..5336
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      5841..5850
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5847..6737
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     polyA_signal    7894..8015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7931..7950)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(8117..8133)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8117..8133)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8130..8152)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8141..8157
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8165..8171,8172..8189,8190..8195))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8210..8231
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8348..8365)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
       61 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      481 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
      601 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
      661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
      721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
      781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
      841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
      901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
      961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1141 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1321 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     1561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     1621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     1681 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     1741 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     1801 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     1861 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     1921 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     1981 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     2041 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     2101 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     2161 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     2221 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     2281 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     2341 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     2401 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     2461 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2521 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2581 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     2641 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
     2701 gactcactat agggagaccc aagcttggta ccgagctcgg atcgatatct gcggcctagc
     2761 tagccaccat ggcttccgtg ctgtcctacg aaagcctggt ccacgccgtg gccggagccg
     2821 tgggaagcgt gacagcaatg acagtgtttt ttcccctgga tacagctaga cttcgacttc
     2881 aggttgatga gaaaagaaaa tccaaaacta cacacatggt gctcctggag atcattaaag
     2941 aagaaggact cctggcacca tatcgagggt ggtttccagt gatttccagt ctctgctgct
     3001 ccaattttgt ctatttctac acttttaata gcctcaaagc actctgggtc aaaggtcaac
     3061 attctaccac tggaaaagat ctggtagttg ggtttgttgc aggagtggtt aatgtgttgc
     3121 taacaactcc actctgggtg gtaaacacca gactgaagct tcaaggagca aaatttagga
     3181 atgaagacat tgtaccaaca aactacaaag gtatcattga tgcttttcat cagatcattc
     3241 gcgatgaagg aatctcggct ttatggaatg gcacatttcc ctcattgctg ttggtcttca
     3301 atcctgccat ccagttcatg ttttatgaag gtttaaaacg gcagctttta aagaaacgga
     3361 tgaagctttc ttccttggat gtgttcatca ttggtgcagt agccaaagcg attgccacca
     3421 cggtgaccta tcccctgcag acggtacagt caattctgag gtttgggcgt catagactaa
     3481 acccagaaaa cagaacattg ggaagtcttc ggaatattct ctatcttctt caccaacgag
     3541 taagacgttt tggaataatg ggactctaca aaggccttga agccaaactg ctgcagacag
     3601 tcctcactgc tgctctcatg ttccttgttt atgagaaact gacagctgcc accttcacag
     3661 ttatggggct gaagcgtgca caccaacacc taccggtcgc caccatggtg agcaagggcg
     3721 aggagggtag cggtggaggt ggatcaggag gtggagttca ggtagaaacc atctccccag
     3781 gagacgggcg caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc
     3841 ttgaagatgg aaagaaattt gattcctccc gggacagaaa caagcccttt aagtttatgc
     3901 taggcaagca ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc
     3961 agagagccaa actgactata tctccagatt atgcctatgg tgccactggg cacccaggca
     4021 tcatcccacc acatgccact ctcgtcttcg atgtggagct tctaaaactg gaagggagtg
     4081 gcagcggagg tagtgggagc ggtggcaagc ccatccccaa ccccctgctg ggcctggaca
     4141 gcaccaaact taatggcatg gacgagcggt acaagtccgg aggaggcggc agcggcggag
     4201 ggggatccat ggagaccgtg agattcggcg gcgacgacat cgagaacagc ctggccaaga
     4261 tggacgacaa ggccctggac aagctggcct tcggcgccat ccagctggac ggcaacggca
     4321 agatcatcca ctacaacgcc gccgagggca ccatcaccgg cagagacccc aagaccgtga
     4381 tcggcaagaa cttcttcacc gacgtggccc ccggcaccca gagcaaggag ttccagggca
     4441 gattcaagga gggcgtgcag aagggcgacc tgaacaccat gttcgagtgg atgatcccca
     4501 ccagcagagg ccccaccaag gtgaaggtgc acatgaagaa ggccatgacc taagcggccg
     4561 catagataac tgatccagtg tgctggaatt aattcgctgt ctgcgagggc cagctgttgg
     4621 ggtgagtact ccctctcaaa agcgggcatg acttctgcgc taagattgtc agtttccaaa
     4681 aacgaggagg atttgatatt cacctggccc gcggtgatgc ctttgagggt ggccgcgtcc
     4741 atctggtcag aaaagacaat ctttttgttg tcaagcttga ggtgtggcag gcttgagatc
     4801 tggccataca cttgagtgac aatgacatcc actttgcctt tctctccaca ggtgtccact
     4861 cccaggtcca actgcaggtc gagcatgcat ctagggcggc caattccgcc cctctccccc
     4921 cccccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     4981 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     5041 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     5101 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     5161 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     5221 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     5281 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     5341 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     5401 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     5461 ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc gggaggcgcg
     5521 ccatgggtga cagctattgg gtctttgtga aacgggtgtc cactagtgaa accgaaaccg
     5581 gaggcggctc cggaggcagc ggcggttctg gaggttccac cctaccggtc tcagctggag
     5641 gcagtgctgg tggaagtgcc ggaggtagaa ttctagaagg tgccgcaaga gaggctgccg
     5701 ctagggaagc agctgcacgt gaggcagctg ctagagaagc tgcagccagg gaggctgcag
     5761 ctagagcatc aggtggtggc ggttcaggcg gaggtggatc tggaggtggc ggatcgggtg
     5821 ggggaggttc gacaccggtc gccaccatgg cagaaatcgg tactggcttt ccattcgacc
     5881 cccattatgt ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg
     5941 gcacccctgt gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca
     6001 tcccgcatgt tgcaccgacc catcgctgca ttgctccaga cctgatcggt atgggcaaat
     6061 ccgacaaacc agacctgggt tatttcttcg acgaccacgt ccgcttcatg gatgccttca
     6121 tcgaagccct gggtctggaa gaggtcgtcc tggtcattca cgactggggc tccgctctgg
     6181 gtttccactg ggccaagcgc aatccagagc gcgtcaaagg tattgcattt atggagttca
     6241 tccgccctat cccgacctgg gacgaatggc cagaatttgc ccgcgagacc ttccaggcct
     6301 tccgcaccac cgacgtcggc cgcaagctga tcatcgatca gaacgttttt atcgagggta
     6361 cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat taccgcgagc
     6421 cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg
     6481 ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata catggactgg ctgcaccagt
     6541 cccctgtccc gaagctgctg ttctggggca ccccaggcgt tctgatccca ccggccgaag
     6601 ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc ccgggtctga
     6661 atctgctgca agaagacaac ccggacctga tcggcagcga gatcgcgcgc tggctgtcga
     6721 cgctcgagat ttccggcatg tacaagtccg gactcagatc tcgagctcaa gcatcgaatt
     6781 ccgaatcccc gcacaaacgc aacctgtact acttctctta cccccttttt gtcgggtttg
     6841 ctctacttcg attggttgct tttcatcttg gattactgtt tgtttggttg tgtcaacgct
     6901 tttctagagc gctgatggct gcgaaacgtt cggctaatag tgaatcgccc cataagagga
     6961 acttatatta cttcagctat cccttgttcg tgggtttcgc gctacttcgg ctggtcgcgt
     7021 ttcatctagg tttgttgttt gtctggttat gtcaacggtt ctcgcgagcg ttaatggcag
     7081 caaagaggtc ggcaaatagc gaatcgcctc ataaacgcaa cctctattac ttctcttacc
     7141 cgttgttcgt tggcttcgct ctccttcggc tagttgcgtt tcatcttggt cttctgttcg
     7201 tgtggctttg ccaaagattt tcgcgagctt taatggcggc taagaggagt gccaatagcg
     7261 aatccccgca taagcggaac ttatactact tctcttaccc tctttttgta ggctttgctc
     7321 tactccggct agtggccttc catctaggcc tactattcgt ctggctctgc cagcgcttct
     7381 cgcgagcctt gatggcagct aagcgttccg caaatagcga atcacctcat aagcggaatc
     7441 tgtactattt ctcatatcct ctcttcgtag ggttcgcact attacgtctt gtagcatttc
     7501 atctgggatt attattcgtc tggttgtgcc agcgattttc acgagcctta atggctgcga
     7561 agagaagcgc aaattctgaa tctcctcaca agcggaacct ctattacttt agttacccgc
     7621 tcttcgtcgg tttcgcgtta ctcaggctgg ttgcattcca cctcggcttg ctgttcgtat
     7681 ggctttgcca gagattctcc cgggcgctaa tggcagccaa gcgctcctaa ggatccaccg
     7741 gatcggtaca agtaactcga gggccaatta attaactcta gataactgat cataatcagc
     7801 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     7861 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     7921 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     7981 agttgtggtt tgtccaaact catcaatgta tcttaacgcg tcgagtgcat tctagttgtg
     8041 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     8101 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     8161 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     8221 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     8281 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     8341 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     8401 ctcactcaaa ggcggtaata cggttatcca cagaatcagg gg
//
