LOCUS       sequence-425782-        5940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lipid droplet localization of CFAST fused to Halotag-6xHp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5940)
  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 5940)
  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: CFAST-Halotag-6xHp
FEATURES             Location/Qualifiers
     source          1..5940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1958..2079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1995..2014)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2049..2068
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2086..2541)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2223..2244)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2435..2454
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2568..2672
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2674..3031
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2674..2865
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2697..2717)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2882..3017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3066..3860
                     /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             3066..3860
                     /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(3120..3139)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3730..3749
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4048..4067)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4092..4139
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4468..5056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4957..4976
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5231..5534
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5535..5738
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5689..5709
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      5765..5774
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5879..5888
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5885..5940,1..835)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
ORIGIN
        1 cgagcgcatg cactacgtcg atgttggtcc gcgcgatggc acccctgtgc tgttcctgca
       61 cggtaacccg acctcctcct acgtgtggcg caacatcatc ccgcatgttg caccgaccca
      121 tcgctgcatt gctccagacc tgatcggtat gggcaaatcc gacaaaccag acctgggtta
      181 tttcttcgac gaccacgtcc gcttcatgga tgccttcatc gaagccctgg gtctggaaga
      241 ggtcgtcctg gtcattcacg actggggctc cgctctgggt ttccactggg ccaagcgcaa
      301 tccagagcgc gtcaaaggta ttgcatttat ggagttcatc cgccctatcc cgacctggga
      361 cgaatggcca gaatttgccc gcgagacctt ccaggccttc cgcaccaccg acgtcggccg
      421 caagctgatc atcgatcaga acgtttttat cgagggtacg ctgccgatgg gtgtcgtccg
      481 cccgctgact gaagtcgaga tggaccatta ccgcgagccg ttcctgaatc ctgttgaccg
      541 cgagccactg tggcgcttcc caaacgagct gccaatcgcc ggtgagccag cgaacatcgt
      601 cgcgctggtc gaagaataca tggactggct gcaccagtcc cctgtcccga agctgctgtt
      661 ctggggcacc ccaggcgttc tgatcccacc ggccgaagcc gctcgcctgg ccaaaagcct
      721 gcctaactgc aaggctgtgg acatcggccc gggtctgaat ctgctgcaag aagacaaccc
      781 ggacctgatc ggcagcgaga tcgcgcgctg gctgtcgacg ctcgagattt ccggcatgta
      841 caagtccgga ctcagatctc gagctcaagc atcgaattcc gaatccccgc acaaacgcaa
      901 cctgtactac ttctcttacc ccctttttgt cgggtttgct ctacttcgat tggttgcttt
      961 tcatcttgga ttactgtttg tttggttgtg tcaacgcttt tctagagcgc tgatggctgc
     1021 gaaacgttcg gctaatagtg aatcgcccca taagaggaac ttatattact tcagctatcc
     1081 cttgttcgtg ggtttcgcgc tacttcggct ggtcgcgttt catctaggtt tgttgtttgt
     1141 ctggttatgt caacggttct cgcgagcgtt aatggcagca aagaggtcgg caaatagcga
     1201 atcgcctcat aaacgcaacc tctattactt ctcttacccg ttgttcgttg gcttcgctct
     1261 ccttcggcta gttgcgtttc atcttggtct tctgttcgtg tggctttgcc aaagattttc
     1321 gcgagcttta atggcggcta agaggagtgc caatagcgaa tccccgcata agcggaactt
     1381 atactacttc tcttaccctc tttttgtagg ctttgctcta ctccggctag tggccttcca
     1441 tctaggccta ctattcgtct ggctctgcca gcgcttctcg cgagccttga tggcagctaa
     1501 gcgttccgca aatagcgaat cacctcataa gcggaatctg tactatttct catatcctct
     1561 cttcgtaggg ttcgcactat tacgtcttgt agcatttcat ctgggattat tattcgtctg
     1621 gttgtgccag cgattttcac gagccttaat ggctgcgaag agaagcgcaa attctgaatc
     1681 tcctcacaag cggaacctct attactttag ttacccgctc ttcgtcggtt tcgcgttact
     1741 caggctggtt gcattccacc tcggcttgct gttcgtatgg ctttgccaga gattctcccg
     1801 ggcgctaatg gcagccaagc gctcctaagg atccaccgga tctagataac tgatcataat
     1861 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1921 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1981 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2041 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     2101 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     2161 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     2221 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2281 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     2341 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2401 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     2461 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     2521 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2581 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2641 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     2701 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2761 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2821 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2881 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2941 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3001 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     3061 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3121 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3181 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3241 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3301 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3361 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3421 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3481 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3541 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     3601 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3661 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3721 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3781 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3841 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     3901 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3961 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4021 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     4081 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     4141 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4201 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4261 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     4321 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4381 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4441 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4501 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4561 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4621 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4681 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4741 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4801 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4861 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4921 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4981 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5041 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5101 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5161 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     5221 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     5281 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     5341 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     5401 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     5461 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     5521 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     5581 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     5641 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     5701 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
     5761 gctagccacc atgggtgaca gctattgggt ctttgtgaaa cgggtgtcca ctagtgaaac
     5821 cgaaaccgga ggcggctccg gaggcagcgg cggttctgga ggttccaccc taccggtcgc
     5881 caccatggca gaaatcggta ctggctttcc attcgacccc cattatgtgg aagtcctggg
//
