LOCUS       sequence-173554-        8565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent protein targeted to lipid droplets (LDs) from the 
            endoplasmic reticulum (ER).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8565)
  AUTHORS   Salo VT, Belevich I, Li S, Karhinen L, Vihinen H, Vigouroux C, Magre
            J, Thiele C, Holtta-Vuori M, Jokitalo E, Ikonen E
  TITLE     Seipin regulates ER-lipid droplet contacts and cargo delivery.
  JOURNAL   EMBO J. 2016 Dec 15;35(24):2699-2716. Epub 2016 Nov 22.
  PUBMED    27879284
REFERENCE   2  (bases 1 to 8565)
  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: "EMBO J."; 
            date: "2016-12-15"; volume: "35(24)"; pages: "2699-2716. Epub 2016 
            Nov 22"
COMMENT     Sequence Label: pEFIRES-P-ACSL3-mCherry
FEATURES             Location/Qualifiers
     source          1..8565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..1178
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          247..1169
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1126..1146
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     intron          1217..1349
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1332..1351
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1394..1412
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3629..3638
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3635..4345
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3780..3798)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4048..4067
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4364..4930
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4380..4932
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4524..4541)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4751..4770
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4932..5534
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     5431..5451
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5768..5902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5805..5824)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5859..5878
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5977..6432
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6064..6083)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6274..6295
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6499..6518)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6618..6640
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6678..6696)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6764..6868
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6869..6937,6938..7729)
                     /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     complement(7087..7106)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7900..8488
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8389..8408
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attcaagctt cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
       61 cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg
      121 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
      181 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
      241 acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct
      301 tgcgtgcctt gaattacttc cacctggctc cagtacgtga ttcttgatcc cgagctggag
      361 ccaggggcgg gccttgcgct ttaggagccc cttcgcctcg tgcttgagtt gaggcctggc
      421 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
      481 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
      541 aagatagtct tgtaaatgcg ggccaggatc tgcacactgg tatttcggtt tttggggccg
      601 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
      661 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
      721 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
      781 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctcc agggggctca aaatggagga
      841 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaggg gcctttccgt
      901 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
      961 agttctggag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     1021 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     1081 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     1141 tggttcaaag tttttttctt ccatttcagg tgtcgtgaac acgggctgca gaagttggtc
     1201 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
     1261 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
     1321 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1381 aggctagagt acttaatacg actcactata ggctagcctc gagaattcta ccatgaataa
     1441 ccacgtgtct tcaaaaccat ctaccatgaa gctaaaacat accatcaacc ctattctttt
     1501 atattttata cattttctaa tatcacttta tactatttta acatacattc cgttttattt
     1561 tttctccgag tcaagacaag aaaaatcaaa ccgaattaaa gcaaagcctg taaattcaaa
     1621 acctgattct gcatacagat ctgttaatag tttggatggt ttggcttcag tattataccc
     1681 tggatgtgat actttagata aagtttttac atatgcaaaa aacaaattta agaacaaaag
     1741 actcttggga acacgtgaag ttttaaatga ggaagatgaa gtacaaccaa atggaaaaat
     1801 ttttaaaaag gttattcttg gacagtataa ttggctttcc tatgaagatg tctttgttcg
     1861 agcctttaat tttggaaatg gattacagat gttgggtcag aaaccaaaga ccaacatcgc
     1921 catcttctgt gagaccaggg ccgagtggat gatagctgca caggcgtgtt ttatgtataa
     1981 ttttcagctt gttacattat atgccactct aggaggtcca gccattgttc atgcattaaa
     2041 tgaaacagag gtgaccaaca tcattactag taaagaactc ttacaaacaa agttgaagga
     2101 tatagtttct ttggtcccac gcctgcggca catcatcact gttgatggaa agccaccgac
     2161 ctggtccgag ttccccaagg gcatcattgt gcataccatg gctgcagtgg aggccctggg
     2221 agccaaggcc agcatggaaa accaacctca tagcaaacca ttgccctcag atattgcagt
     2281 aatcatgtac acaagtggat ccacaggact tccaaaggga gtcatgatct cacatagtaa
     2341 cattattgct ggtataactg ggatggcaga aaggattcca gaactaggag aggaagatgt
     2401 ctacattgga tatttgcctc tggcccatgt tctagaatta agtgctgagc ttgtctgtct
     2461 ttctcacgga tgccgcattg gttactcttc accacagact ttagcagatc agtcttcaaa
     2521 aattaaaaaa ggaagcaaag gggatacatc catgttgaaa ccaacactga tggcagcagt
     2581 tccggaaatc atggatcgga tctacaaaaa tgtcatgaat aaagtcagtg aaatgagtag
     2641 ttttcaacgt aatctgttta ttctggccta taattacaaa atggaacaga tttcaaaagg
     2701 acgtaatact ccactgtgcg acagctttgt tttccggaaa gttcgaagct tgctaggggg
     2761 aaatattcgt ctcctgttgt gtggtggcgc tccactttct gcaaccacgc agcgattcat
     2821 gaacatctgt ttctgctgtc ctgttggtca gggatacggg ctcactgaat ctgctggggc
     2881 tggaacaatt tccgaagtgt gggactacaa tactggcaga gtgggagcac cattagtttg
     2941 ctgtgaaatc aaattaaaaa actgggagga aggtggatac tttaatactg ataagccaca
     3001 ccccaggggt gaaattctta ttgggggcca aagtgtgaca atggggtact acaaaaatga
     3061 agcaaaaaca aaagctgatt tctttgaaga tgaaaatgga caaaggtggc tctgtactgg
     3121 ggatattgga gagtttgaac ccgatggatg cttaaagatt attgatcgta aaaaggacct
     3181 tgtaaaacta caggcagggg aatatgtttc tcttgggaaa gtagaggcag ctttgaagaa
     3241 tcttccacta gtagataaca tttgtgcata tgcaaacagt tatcattctt atgtcattgg
     3301 atttgttgtg ccaaatcaaa aggaactaac tgaactagct cgaaagaaag gacttaaagg
     3361 gacttgggag gagctgtgta acagttgtga aatggaaaat gaggtactta aagtgctttc
     3421 cgaagctgct atttcagcaa gtctggaaaa gtttgaaatt ccagtaaaaa ttcgtttgag
     3481 tcctgaaccg tggacccctg aaactggtct ggtgacagat gccttcaagc tgaaacgcaa
     3541 agagcttaaa acacattacc aggcggacat tgagcgaatg tatggaagaa aacagtcgac
     3601 ggtaccgcgg gcccgggatc caccggtcgc caccatggtg agcaagggcg aggaggataa
     3661 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     3721 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     3781 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     3841 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     3901 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     3961 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     4021 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     4081 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     4141 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     4201 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     4261 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     4321 cggcatggac gagctgtaca agtagcggcc gctctagccc aattccgccc ccccccctaa
     4381 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     4441 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     4501 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     4561 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     4621 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     4681 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     4741 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     4801 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     4861 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     4921 acgatgataa tatgggcacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     4981 tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     5041 ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     5101 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     5161 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     5221 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     5281 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     5341 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     5401 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     5461 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     5521 agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac
     5581 cccatggctc cgaccgaagc cacccggggc ggccccgccg accccgcacc cgcccccgag
     5641 gcccaccgac tctagctaga ggatcataat cagccatacc acatttgtag aggttttact
     5701 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     5761 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5821 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5881 tgtatcttat catgtctgga tccgggctgg cgtaatagcg aagaggcccg caccgatcgc
     5941 ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa
     6001 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     6061 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     6121 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     6181 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     6241 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     6301 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     6361 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     6421 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     6481 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     6541 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     6601 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     6661 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     6721 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6781 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6841 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6901 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6961 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     7021 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     7081 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     7141 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     7201 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     7261 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     7321 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     7381 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     7441 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7501 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7561 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     7621 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7681 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7741 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7801 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7861 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7921 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7981 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     8041 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     8101 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     8161 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     8221 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     8281 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     8341 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     8401 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     8461 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     8521 ggccttttgc tggccttttg ctcacatggc tcgacagatc cccca
//
