LOCUS Exported 5400 bp ds-DNA circular SYN 22-DEC-2022 DEFINITION Homology arms and mEGFP-SGLRS for N-terminus tagging of human G3BP stress granule assembly factor 1. ACCESSION . VERSION . KEYWORDS AICSDP-078:G3BP1-mEGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5400) TITLE Allen Institute for Cell Science Plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 5400) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 22, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5400 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(47..69) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 169..188 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 382..404 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 396..413 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 397..413 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 1431..2147 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1476..1497) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1737..1756) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2084..2105 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(3197..3213) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3197..3213) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3210..3232) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3221..3237 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(3245..3275) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3290..3311 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3428..3445) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3599..4187) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3679..3698) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4358..5218) /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 4981..5000 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5219..5323) /gene="bla" /label=AmpR promoter primer_bind 5391..9 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 61 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 121 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc 181 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 241 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 301 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa 361 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattga 421 cgcgtattgg gatattggac ttcttatggg acaggtgcct ttcttggcat tcaaaggtgt 481 taattttaca tgatcacaat tttgatacag taggaattat tttgagtata aacatcttgt 541 ttgttaactt ccccatttta ttttgaaatt ttttaattac atgatagaat gaagtagtaa 601 gcatatttag aatttttagt cttgtagttt ctttttaaaa gatgctatac taaaagttta 661 tctgatttag atgggccaga aaatgtgata gtttgtcccc ggatgtcaaa actatgttgt 721 ttaagattat tgcctttgaa tctaatattg cctcaaataa gaatgtgctt gtaactttac 781 aactggaaca ctcttaattg ggcttaataa cttaagaaaa ttaccaggtg ctgtggctca 841 cgcctgtaat cccagcactt tgggaggctg aggcgggcag atcatgaggt caggagtttg 901 agaccagcct ggataacata gtgaaatccc gtgctaaaaa atacaaaaat tagccgggca 961 tgatggcgct tgccagtagt cccagctact atggaggctg aggcaggaga atcgcttgaa 1021 cctgggaggt ggaggttgtg gtgagccgag atcgcaccac tgcactccag cctgggcaac 1081 agagagactc tgtctcaaaa acaaaacaag aaaattatgg tgggaagtat gttggaaata 1141 tatacctttg aggttcttac gtcttgaata ttggggataa tgatgaattc ttcataatga 1201 tttccccctt ttgtattcat atgattgcaa tacatttgta taaaatctca actggtctgt 1261 atgggctctg tttgttttta tgatgtttgt tcatgtttta aacgacttgc tgaaatgatc 1321 ttgtctctga gttggtttca gctaaatgat tcggtctttt cccctgtgtt tgatccttca 1381 ggtttggaca tatttgactc ttttccccgc aggttgaatt gaccaaagca atggtgagca 1441 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa 1501 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga 1561 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca 1621 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact 1681 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg 1741 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca 1801 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt 1861 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg 1921 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc 1981 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca 2041 cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt 2101 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagagc ggcctgagaa 2161 gcgtgatgga gaagccttct cccctcctcg tcgggcggga atttgtgaga cagtattaca 2221 cactgctgaa ccaggcccca gacatgctgc ataggtaaga catttttctc ctgcatcatc 2281 taatgctgtc ttttagtatg tggtacttat ctttgagatt ctcccactca tcatcttatg 2341 ctttgtaggg ttgttgcata atacttattt acatataaat atcccccaat tttttatttg 2401 agacagagtc tcgccctgtt gtccagactg gagtgcagtg gtgcgatctg gactgactgc 2461 aacgtccacc tccccagttt aagtgattct tctgccccag cctcctgagt agctgggacc 2521 acaggtgccc acaaccatgc ccggctaatt tttgtatttt tttttttttt ttttttaagt 2581 agagacgggg tttcaccatg ttggccaggc tggtctcaaa ctcctaacct caggtgattg 2641 atccgccttg gcctcccaaa gtcctgggat tacaggcgtg agccactgcg cctagcccca 2701 aaaaataatt tttgaaagtt aaactaagtt atctaaactt gtacattatt ggaaaataac 2761 aattgaaatg taaaggtata gtcatgcatt atttactaag aatgtattaa agattgctgg 2821 agctgaggtg gatgtaaaaa ataagaatgt attgaagaca atccatgact gttcctgtaa 2881 taccatactt ttgtgttgat gcgctttaaa aaattagtgt ctttagtaga aatgatgcat 2941 gcttattgaa aattagaaca cagataaatt atgaaagtag taaaaccaca gaaacaattt 3001 ttctatatat atagaaacat gttttaatat actgtcttaa ctttcttact tgataatctg 3061 gagtcatctt tcagggtaat acttgtatcc cattaatttt agtggctaca cagcttttta 3121 tagagatgta gtgtacttta atctcctaat gatggatcta taatcccaat ggcgcgccga 3181 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 3241 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 3301 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 3361 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 3421 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 3481 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 3541 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 3601 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 3661 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 3721 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 3781 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 3841 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 3901 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 3961 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 4021 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 4081 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 4141 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 4201 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 4261 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 4321 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 4381 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 4441 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 4501 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 4561 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 4621 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 4681 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 4741 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 4801 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 4861 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 4921 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 4981 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 5041 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 5101 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 5161 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 5221 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 5281 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 5341 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta //