LOCUS Exported 7756 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Fluorescent protein localized to lipid droplets (LDs). ACCESSION . VERSION . KEYWORDS pEFIRES-P-EGFP-ADRP 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7756 /organism="synthetic DNA construct" /mol_type="other DNA" 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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1434..2150 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1434..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 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 6060..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 //