LOCUS Exported 4935 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION M11-92 expression. ACCESSION . VERSION . KEYWORDS DsRed2-M11-92 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4935) AUTHORS Chang CL, Weigel AV, Ioannou MS, Pasolli HA, Xu CS, Peale DR, Shtengel G, Freeman M, Hess HF, Blackstone C, Lippincott-Schwartz J TITLE Spastin tethers lipid droplets to peroxisomes and directs fatty acid trafficking through ESCRT-III. JOURNAL J Cell Biol. 2019 Aug 5;218(8):2583-2599. doi: 10.1083/jcb.201902061. Epub 2019 Jun 21. PUBMED 31227594 REFERENCE 2 (bases 1 to 4935) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4935 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 16..319 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 320..523 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 474..494 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 562..571 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 568..1242 /codon_start=1 /product="improved tetrameric variant of DsRed fluorescent protein" /label=DsRed2 /note="mammalian codon-optimized" /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH LFL" primer_bind complement(748..768) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 1160..1183 /label=DsRed1-C /note="DsRed1, forward primer" polyA_signal 1678..1799 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1715..1734) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1769..1788 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1806..2261) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1943..1964) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2155..2174 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2288..2392 /gene="bla" /label=AmpR promoter promoter 2394..2751 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2417..2437) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2602..2737 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2664..2683 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2786..3580 /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(2840..2859) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3450..3469 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3768..3787) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3812..3859 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4188..4776 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4677..4696 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca 541 gatccgctag cgctaccggt cgccaccatg gcctcctccg agaacgtcat caccgagttc 601 atgcgcttca aggtgcgcat ggagggcacc gtgaacggcc acgagttcga gatcgagggc 661 gagggcgagg gccgccccta cgagggccac aacaccgtga agctgaaggt gaccaagggc 721 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg 781 tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc 841 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg cgaccgtgac ccaggactcc 901 tccctgcagg acggctgctt catctacaag gtgaagttca tcggcgtgaa cttcccctcc 961 gacggccccg tgatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac 1021 ccccgcgacg gcgtgctgaa gggcgagacc cacaaggccc tgaagctgaa ggacggcggc 1081 cactacctgg tggagttcaa gtccatctac atggccaaga agcccgtgca gctgcccggc 1141 tactactacg tggacgccaa gctggacatc acctcccaca acgaggacta caccatcgtg 1201 gagcagtacg agcgcaccga gggccgccac cacctgttcc tgtccggact cagatctcga 1261 gctcaagctt cgaattctcc gggtggacga gggaagaaga aaggctccgg cggcgccagc 1321 aacccggtgc ctcccaggcc tccgccccct tgcctggccc ccgcccctcc cgccgccggg 1381 ccggcccctc cgcccgagtc gccgcataag cggaacctgt actatttctc ctacccgctg 1441 tttgtaggct tcgcgctgct gcgtttggtc gccttccacc tggggctcct cttcgtgtgg 1501 ctctgccagc gcttctcccg cgccctcatg gcagccaaga ggagctaagg atccaccgga 1561 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa 1621 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac 1681 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 1741 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa 1801 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag 1861 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac 1921 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga 1981 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc 2041 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg 2101 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa 2161 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac 2221 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg 2281 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct 2341 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc 2401 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca 2461 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc 2521 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata 2581 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 2641 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 2701 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca 2761 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc 2821 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 2881 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 2941 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac 3001 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 3061 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 3121 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 3181 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 3241 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 3301 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 3361 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 3421 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 3481 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 3541 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa 3601 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 3661 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 3721 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag 3781 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac 3841 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat 3901 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac 3961 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc 4021 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 4081 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt 4141 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 4201 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 4261 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 4321 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta 4381 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 4441 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 4501 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 4561 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 4621 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 4681 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 4741 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 4801 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat 4861 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc 4921 attagttcat agccc //