LOCUS       sequence-173556-        6559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent marker of lipid droplets (LDs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6559)
  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 6559)
  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-mCherry-HPos
FEATURES             Location/Qualifiers
     source          1..6559
                     /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      1428..1437
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1434..2141
                     /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(1579..1597)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1847..1866
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2358..2924
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2374..2926
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2518..2535)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2745..2764
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2926..3528
                     /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     3425..3445
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3762..3896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3799..3818)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3853..3872
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3971..4426
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4058..4077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4268..4289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4493..4512)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4612..4634
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4672..4690)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4758..4862
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4863..4931,4932..5723)
                     /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(5081..5100)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5894..6482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6383..6402
                     /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 ggctagcgct accggtcgcc accatggtga
     1441 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     1501 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     1561 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     1621 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     1681 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     1741 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     1801 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     1861 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     1921 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     1981 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     2041 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     2101 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtccggaatg gatgtcctgg
     2161 ttccattgct gcagctgctg gtgctgctcc tgactctgcc tttacacctg ctggctctgc
     2221 tgggctgctg gcagcccctc tttgaagcga ttggcaagat attcagcaat atccgcatca
     2281 gcacgcagaa agagatatga gggatcattc acgcgtggta cctctagagt cgacccgggc
     2341 ggccgctcta gcccaattcc gccccccccc ctaacgttac tggccgaagc cgcttggaat
     2401 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     2461 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     2521 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     2581 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     2641 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     2701 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     2761 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     2821 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     2881 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggg caccgagtac
     2941 aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc
     3001 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag
     3061 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     3121 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     3181 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     3241 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     3301 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     3361 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     3421 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     3481 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
     3541 gacccgcagc gcccgaccga aaggagcgca cgaccccatg gctccgaccg aagccacccg
     3601 gggcggcccc gccgaccccg cacccgcccc cgaggcccac cgactctagc tagaggatca
     3661 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3721 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3781 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3841 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccggg
     3901 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3961 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4021 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4081 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4141 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4201 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4261 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4321 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     4381 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     4441 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     4501 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4561 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4621 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4681 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4741 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4801 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4861 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4921 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4981 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5041 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5101 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5161 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5221 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5281 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5341 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5401 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5461 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5521 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5581 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5641 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5701 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5761 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5821 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5881 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5941 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6001 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6061 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6121 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6181 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6241 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6301 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6361 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6421 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6481 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6541 tggctcgaca gatccccca
//
