LOCUS       sequence-427381-        4800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ER marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4800)
  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 4800)
  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: mApple-ERM
FEATURES             Location/Qualifiers
     source          1..4800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..48
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(66..521)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(203..224)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     415..434
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        548..652
                     /gene="bla"
                     /label=AmpR promoter
     promoter        654..1011
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        654..845
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(677..697)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      862..997
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1046..1840
                     /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             1046..1840
                     /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(1100..1119)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1710..1729
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2028..2047)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2072..2119
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2448..3036
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2937..2956
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3211..3514
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3515..3718
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3669..3689
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3757..3766
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3763..4470
                     /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     4176..4195
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    join(4738..4800,1..59)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4775..4794)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
       61 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
      121 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      181 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      241 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      301 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      361 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      421 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      481 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
      541 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
      601 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
      661 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
      721 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
      781 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
      841 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
      901 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
      961 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     1021 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     1081 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     1141 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     1201 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     1261 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     1321 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     1381 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     1441 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     1501 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     1561 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     1621 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     1681 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     1741 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     1801 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     1861 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     1921 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     1981 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     2041 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     2101 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     2161 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     2221 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     2281 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2341 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     2401 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2461 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2521 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2581 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2641 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2701 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2761 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2821 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2881 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2941 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3001 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3061 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3121 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     3181 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     3241 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     3301 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     3361 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     3421 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     3481 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     3541 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     3601 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     3661 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     3721 tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca ccatggtgag caagggcgag
     3781 gagaataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     3841 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgaggccttt
     3901 cagaccgcta agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     3961 tcccctcagt tcatgtacgg ctccaaggtc tacattaagc acccagccga catccccgac
     4021 tacttcaagc tgtccttccc cgagggcttc aggtgggagc gcgtgatgaa cttcgaggac
     4081 ggcggcatta ttcacgttaa ccaggactcc tccctgcagg acggcgtgtt catctacaag
     4141 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     4201 ggctgggagg cctccgagga gcggatgtac cccgaggacg gcgccctgaa gagcgagatc
     4261 aagaagaggc tgaagctgaa ggacggcggc cactacgccg ccgaggtcaa gaccacctac
     4321 aaggccaaga agcccgtgca gctgcccggc gcctacatcg tcgacatcaa gttggacatc
     4381 gtgtcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     4441 tccaccggcg gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcg
     4501 atcaccaccg tggagtccaa ctcctcctgg tggaccaact gggtgatccc cgccatctcc
     4561 gccctggtgg tggccctgat gtaccgcctg tacatggccg aggactaagg atccaccgga
     4621 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     4681 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     4741 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
//
