LOCUS       sequence-173553-        7756 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Fluorescent protein localized to lipid droplets  (LDs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7756)
  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 7756)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-EGFP-ADRP
FEATURES             Location/Qualifiers
     source          1..7756
                     /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             join(1434..1436,1437..1439,1440..2150)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1434..1436,1437..1439,1440..2150)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1479..1500)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1740..1759)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2087..2108
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3555..4121
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3571..4123
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3715..3732)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3942..3961
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4123..4725
                     /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     4622..4642
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4959..5093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4996..5015)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5050..5069
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5168..5623
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5255..5274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5465..5486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5690..5709)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5809..5831
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5869..5887)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5955..6059
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6060..6128,6129..6920)
                     /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(6278..6297)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7091..7679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7580..7599
                     /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 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     1501 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     1561 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1621 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     1681 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1741 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1801 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1861 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     1921 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     1981 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     2041 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     2101 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca
     2161 gatctcgagc tatggcatcc gttgcagttg atccacaacc gagtgtggtg actcgggtgg
     2221 tcaacctgcc cttggtgagc tccacgtatg acctcatgtc ctcagcctat ctcagtacaa
     2281 aggaccagta tccctacctg aagtctgtgt gtgagatggc agagaacggt gtgaagacca
     2341 tcacctccgt ggccatgacc agtgctctgc ccatcatcca gaagctagag ccgcaaattg
     2401 cagttgccaa tacctatgcc tgtaaggggc tagacaggat tgaggagaga ctgcctattc
     2461 tgaatcagcc atcaactcag attgttgcca atgccaaagg cgctgtgact ggggcaaaag
     2521 atgctgtgac gactactgtg actggggcca aggattctgt ggccagcacg atcacagggg
     2581 tgatggacaa gaccaaaggg gcagtgactg gcagtgtgga gaagaccaag tctgtggtca
     2641 gtggcagcat taacacagtc ttggggagtc ggatgatgca gctcgtgagc agtggcgtag
     2701 aaaatgcact caccaaatca gagctgttgg tagaacagta cctccctctc actgaggaag
     2761 aactagaaaa agaagcaaaa aaagttgaag gatttgatct ggttcagaag ccaagttatt
     2821 atgttagact gggatccctg tctaccaagc ttcactcccg tgcctaccag caggctctca
     2881 gcagggttaa agaagctaag caaaaaagcc aacagaccat ttctcagctc cattctactg
     2941 ttcacctgat tgaatttgcc aggaagaatg tgtatagtgc caatcagaaa attcaggatg
     3001 ctcaggataa gctctacctc tcatgggtag agtggaaaag gagcattgga tatgatgata
     3061 ctgatgagtc ccactgtgct gagcacattg agtcacgtac tcttgcaatt gcccgcaacc
     3121 tgactcagca gctccagacc acgtgccaca ccctcctgtc caacatccaa ggtgtaccac
     3181 agaacatcca agatcaagcc aagcacatgg gggtgatggc aggcgacatc tactcagtgt
     3241 tccgcaatgc tgcctccttt aaagaagtgt ctgacagcct cctcacttct agcaaggggc
     3301 agctgcagaa aatgaaggaa tctttagatg acgtgatgga ttatcttgtt aacaacacgc
     3361 ccctcaactg gctggtaggt cccttttatc ctcagctgac tgagtctcag aatgctcagg
     3421 accaaggtgc agagatggac aagagcagcc aggagaccca gcgatctgag cataaaactc
     3481 attaagtcga gctcaagctt cgaattcacg cgtggtacct ctagagtcga cccgggcggc
     3541 cgctctagcc caattccgcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     3601 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     3661 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     3721 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     3781 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     3841 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     3901 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     3961 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     4021 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     4081 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatgggcac cgagtacaag
     4141 cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc
     4201 gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg
     4261 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     4321 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4381 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4441 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     4501 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4561 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4621 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4681 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac
     4741 ccgcagcgcc cgaccgaaag gagcgcacga ccccatggct ccgaccgaag ccacccgggg
     4801 cggccccgcc gaccccgcac ccgcccccga ggcccaccga ctctagctag aggatcataa
     4861 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     4921 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     4981 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5041 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgggctg
     5101 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     5161 cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5221 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5281 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5341 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5401 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5461 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5521 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5581 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     5641 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
     5701 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     5761 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     5821 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     5881 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5941 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6001 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6061 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6121 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6181 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6241 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6301 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6361 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6421 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6481 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6541 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6601 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6661 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6721 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6781 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6841 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6901 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6961 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7021 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7081 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7141 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7201 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     7261 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7321 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7381 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7441 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7501 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7561 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7621 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7681 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg
     7741 ctcgacagat ccccca
//
