LOCUS Exported 4647 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses FRB-NFAST in mammalian cells. ACCESSION . VERSION . KEYWORDS pAG148 FRB-NFAST SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4647) AUTHORS Tebo AG, Gautier A TITLE A split fluorescent reporter with rapid and reversible complementation. JOURNAL Nat Commun. 2019 Jun 27;10(1):2822. doi: 10.1038/s41467-019-10855-0. PUBMED 31249300 REFERENCE 2 (bases 1 to 4647) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4647 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 218..238 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter complement(263..367) /gene="bla" /label=AmpR promoter rep_origin 394..849 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(481..500) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 691..712 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 856..977 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(867..886) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 921..940 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(1483..1752) /codon_start=1 /product="FKBP-rapamycin binding domain of human FRAP" /label=FRB /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ" CDS complement(1759..1788) /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" regulatory 1788..1797 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(1844..2047) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(1873..1893) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 2048..2351 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" rep_origin complement(2526..3114) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2606..2625) /label=pBR322ori-F /note="pBR322 origin, forward primer" polyA_signal 3443..3490 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" primer_bind 3515..3534 /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" CDS complement(3722..4516) /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(3833..3852) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 4443..4462 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(4551..261) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4565..53 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(4619..4638) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" ORIGIN 1 agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga 61 gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct 121 gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc 181 atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac 241 agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc atttatcagg 301 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 361 ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg 421 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 481 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 541 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 601 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 661 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac 721 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt 781 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc 841 ttacaattta cgccttaaga tacattgatg agtttggaca aaccacaact agaatgcagt 901 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa 961 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg 1021 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg 1081 atctagagtc gcggccgcct attaggaaag ggctttcttc atgtgcacct tgaccttggt 1141 tggtcccctg cttgtcggta tcatccattc gaacatggtg ttcagattcc ctgacgctac 1201 gccttccttg aatttgccgt aaaactcggg agaatccgtt ccaggtgcaa catccttgaa 1261 gaagttcttc ccaatcacct gtttgggatc tctgcctgtg atgtctcctt cagcagcatt 1321 gtactgcagg atattcccgt caccatcgag ctgaattgcg ccaaaggcca acccatccag 1381 ttgtccgtcg tccattttgg ccagagtgtt ctcgatgtcc tcactgccaa aggcaacatg 1441 ctccatggat ccccctccgc cgctgccgcc tcctccggag acctgctttg agattcgtcg 1501 gaacacatga taatagaggt cccaggcttg ggtgaggtcc ttgacattcc ctgatttcat 1561 gtacttcctg caccactctt gggcctccat taaatctcga ccataggcct gattaaagga 1621 tgtttccttc agagtctggg ggccccgttc catcatagca tgcaagggct ccagcacctc 1681 aaacatgcct ttcacgttcc tttccccaaa gtacaaacga gatgcctctt ccaggccttc 1741 atgccacatc tcgaattcca agtcctcttc agaaataagc ttttgttcca tggtggcaga 1801 tctgagtccg gtagcgctag cggatctgac ggttcactaa accagctctg cttatataga 1861 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt 1921 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga 1981 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc 2041 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc 2101 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag 2161 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat 2221 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca 2281 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa 2341 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact 2401 attaataact aatgcatggc ggtaatacgg ttatccacag aatcagggga taacgcagga 2461 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 2521 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 2581 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 2641 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 2701 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 2761 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 2821 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 2881 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 2941 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 3001 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 3061 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 3121 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 3181 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 3241 tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg 3301 acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa 3361 gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc 3421 tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt 3481 tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc 3541 ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag 3601 ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg 3661 aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg 3721 ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga 3781 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac 3841 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga 3901 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca 3961 cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg 4021 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag 4081 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa 4141 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt 4201 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt 4261 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc 4321 gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa 4381 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt 4441 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc 4501 catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag 4561 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc 4621 tcggcctctg cataaataaa aaaaatt //