LOCUS Exported 7017 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses myc-dL5(E52D)-mCer3 fusion protein in nuclei of mammalian cells. (MBIC5, dL5**, FAP). ACCESSION . VERSION . KEYWORDS pcDNA3.1-NLS-myc-dL5-2xG4S-mCer3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7017) AUTHORS Telmer CA, Verma R, Teng H, Andreko S, Law L, Bruchez MP TITLE Rapid, specific, no-wash, far-red fluorogen activation in subcellular compartments by targeted fluorogen activating proteins. JOURNAL ACS Chem Biol. 2015 May 15;10(5):1239-46. doi: 10.1021/cb500957k. Epub 2015 Feb 16. PUBMED 25650487 REFERENCE 2 (bases 1 to 7017) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7017 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 619..822 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 773..793 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 867..886 /label=T7 /note="T7 promoter, forward primer" promoter 867..885 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 1011..1020 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1020..1049 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1065..1784 /codon_start=1 /product="fluorogen-activating protein composed of a dimer of the L5-MG single-chain antibody fragment (Szent-Gyorgyi et al., 2008)" /label=FAP-alpha-2 /note="binds malachite green derivatives to produce red fluorescence" /translation="QAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGHYANWFQQKPGQA PRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSLSDVDGYLFGGGT QLTVLSGGGGSGGGGSGGGGSGGGGSQAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGH YANWFQQKPGQAPRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSL SDVDGYLFGGGTQLTVLS" primer_bind complement(1887..1908) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2148..2167) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2495..2516 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(2611..2628) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2617..2841 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2887..3315 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2974..2993) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3184..3205 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3324..3344) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3329..3658 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3509..3644 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3571..3590 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3725..4519 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3779..3798) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4389..4408 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4693..4814 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4730..4749) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4784..4803 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4863..4879) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4863..4879) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4876..4898) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4887..4903 /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(4911..4941) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4956..4977 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5094..5111) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5265..5850) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5345..5364) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6021..6881) /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 6644..6663 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6882..6986) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg 541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta 601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg 661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg 721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg 781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa 841 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagctggc 901 tagcgtttaa acttaagctt gccaccggat ccgccaccat gaacaagaac tctgctaagc 961 gtcgtaagaa gggtacttct gctaagacta aacgtcctaa agttgaattc gccaccatgg 1021 agcagaagct gatcagcgag gaggacctgg gcctttctgg cctgcaggcc gtcgttaccc 1081 aagaacctag tgttaccgtt agcccaggtg gtactgttat acttacttgt ggaagtggta 1141 cgggtgccgt cacatctggt cattatgcaa attggtttca acaaaaacca ggacaagctc 1201 caagagcttt gatttttgat actgataaga agtattcttg gaccccaggt agattttctg 1261 gatctttgct gggagcaaag gcagctttga caatatcaga tgctcagcct gaggacgaag 1321 ccgagtatta ctgttctctt agcgacgtgg atggctactt gtttggcggt ggaacacaac 1381 tgacggttct gtccggtggt ggcggctctg gtggcggtgg cagcggcggt ggtggttccg 1441 gaggcggcgg ttctcaggct gtggtgactc aggagccgtc agtgactgtg tccccaggag 1501 ggacagtcat tctcacttgt ggctccggca ctggagctgt caccagtggt cattatgcca 1561 actggttcca gcagaagcct ggccaagccc ccagggcact tatatttgac accgacaaga 1621 agtattcctg gacccctggc cgattctcag gctccctcct tggggccaag gctgccctga 1681 ccatctcgga tgcgcagcct gaagatgagg ctgagtatta ctgttcgctc tccgacgttg 1741 acggttatct gttcggagga ggcacccagc tgaccgtcct ctccgcggcc ggtctggccc 1801 atatgggagg cggtgggtct ggtggcggtg gcagcaccgg tatggtgagc aagggcgagg 1861 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca 1921 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt 1981 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgagct 2041 ggggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt 2101 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact 2161 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga 2221 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacgcca 2281 tccacggcaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttcg 2341 gcctcaactg caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca 2401 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca 2461 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg 2521 ccgccgggat cactctcggc atggacgagc tgtacaagta actcgaggca gctgtcgact 2581 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat 2641 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc 2701 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg 2761 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg 2821 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt 2881 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 2941 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 3001 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 3061 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 3121 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 3181 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg 3241 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa 3301 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg 3361 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg 3421 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 3481 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca 3541 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc 3601 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa 3661 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt 3721 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct 3781 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct 3841 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga 3901 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc 3961 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg 4021 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc 4081 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca 4141 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga 4201 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc 4261 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga 4321 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca 4381 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg 4441 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct 4501 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc 4561 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 4621 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 4681 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 4741 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 4801 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 4861 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 4921 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 4981 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 5041 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 5101 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 5161 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 5221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 5281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 5341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 5401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 5461 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 5521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 5581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 5641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 5701 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 5761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag 5821 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 5881 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 5941 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 6001 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 6061 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 6121 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 6181 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 6241 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 6301 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 6361 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 6421 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 6481 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 6541 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 6601 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 6661 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 6721 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 6781 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 6841 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 6901 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 6961 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //