LOCUS Exported 7062 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses myc-dL5(E52D)-mCer3 fusion protein in endoplasmic reticulum of mammalian cells with an Igk-leader. (MBIC5, dL5**, FAP). ACCESSION . VERSION . KEYWORDS pcDNA3.1-kappa-myc-dL5-2XG4S-mCer3-KDEL SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7062) 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 7062) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7062 /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" sig_peptide 951..1013 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader 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(2656..2673) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2662..2886 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2932..3360 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3019..3038) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3229..3250 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3369..3389) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3374..3703 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3554..3689 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3616..3635 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3770..4564 /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(3824..3843) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4434..4453 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4738..4859 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4775..4794) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4829..4848 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4908..4924) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4908..4924) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4921..4943) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4932..4948 /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(4956..4986) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5001..5022 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5139..5156) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5310..5895) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5390..5409) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6066..6926) /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 6689..6708 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6927..7031) /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 ccggcttggg gatatccacc atggagacag 961 acacactcct gctatgggta ctgctgctct gggttccagg ttccactggt gacgaattcg 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 tgtacaagct cgagaaggat gaactgtaat 2581 gattatcaca actcgtcttt ctcgaggtcg aggcagctgt cgactctaga gggcccgttt 2641 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct 2701 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg 2761 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc 2821 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct 2881 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct 2941 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 3001 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 3061 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 3121 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 3181 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 3241 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 3301 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 3361 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 3421 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 3481 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 3541 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 3601 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca 3661 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg 3721 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca 3781 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 3841 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 3901 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc 3961 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 4021 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 4081 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 4141 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 4201 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 4261 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct 4321 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 4381 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 4441 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 4501 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 4561 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga 4621 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 4681 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac 4741 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 4801 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 4861 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 4921 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 4981 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 5041 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 5101 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 5161 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 5221 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 5281 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 5341 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 5401 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 5461 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 5521 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 5581 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 5641 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 5701 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 5761 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 5821 ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga 5881 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 5941 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 6001 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 6061 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 6121 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 6181 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 6241 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 6301 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 6361 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 6421 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 6481 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 6541 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 6601 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 6661 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 6721 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 6781 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 6841 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 6901 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 6961 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 7021 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc //