LOCUS Exported 5428 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Measure FRET between LSSmOrange and mKate2. ACCESSION . VERSION . KEYWORDS pLSSmOrange-mKate2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5428) AUTHORS Demeautis C, Sipieter F, Roul J, Chapuis C, Padilla-Parra S, Riquet FB, Tramier M TITLE Multiplexing PKA and ERK1 JOURNAL Sci Rep. 2017 Jan 20;7:41026. doi: 10.1038/srep41026. PUBMED 28106114 REFERENCE 2 (bases 1 to 5428) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5428 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 169..472 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 473..676 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 627..647 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 715..724 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 721..1428 /codon_start=1 /product="monomeric orange fluorescent protein with a large Stokes shift (Shcherbakova et al., 2012)" /label=LSSmOrange /note="mammalian codon-optimized" /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind 1134..1153 /label=mCherry-F /note="mCherry, forward primer" misc_feature 1434..1490 /label=MCS /note="multiple cloning site" regulatory 1498..1507 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1504..2202 /codon_start=1 /product="monomeric far-red fluorescent protein (Shcherbo et al., 2009)" /label=mKate2 /note="mammalian codon-optimized" /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY CDLPSKLGHR" polyA_signal 2324..2445 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2361..2380) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2415..2434 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2452..2907) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2589..2610) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2801..2820 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2934..3038 /gene="bla" /label=AmpR promoter promoter 3040..3397 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3063..3083) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3248..3383 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3310..3329 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3432..4226 /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(3486..3505) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4096..4115 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4414..4433) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4458..4505 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4834..5422 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5323..5342 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 61 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata 121 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact 181 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat 241 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta 301 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc 361 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg 421 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg 481 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct 541 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa 601 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt 661 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc 721 atggtgagca agggcgagga gaataacatg gccatcatca aggagttcat gcgcttcaag 781 gtgcgcatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc 841 cgcccctacg agggctttca gaccgttaag ctgaaggtga ccaagggtgg ccccctgccc 901 ttcgcctggg acatcttgtc ccctcagttc acctacggct ccaaggccta cgtgaagcac 961 cccgccgaca tccccgacta cctcaagctg tccttccccg agggcttcaa gtgggagcgc 1021 gtgatgaact tcgaggacgg cggcgtggtg accgtgactc aggactcctc cctgcaggac 1081 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta 1141 atgcagaaga agaccatggg catggaggcc tcctccgagc ggatgtaccc cgaggacggc 1201 gccctgaagg gcgaggacaa gctcaggctg aagctgaagg acggcggcca ctacacctcc 1261 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagt tgcccggcgc ctacatcgtc 1321 gacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa 1381 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta ctcagatctc 1441 gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc 1501 accatggtga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc 1561 gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc 1621 cagaccatga gaatcaaggc ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg 1681 gctaccagct tcatgtacgg cagcaaaacc ttcatcaacc acacccaggg catccccgac 1741 ttctttaagc agtccttccc cgagggcttc acatgggaga gagtcaccac atacgaagac 1801 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac 1861 gtcaagatca gaggggtgaa cttcccatcc aacggccctg tgatgcagaa gaaaacactc 1921 ggctgggagg cctccaccga gaccctgtac cccgctgacg gcggcctgga aggcagagcc 1981 gacatggccc tgaagctcgt gggcgggggc cacctgatct gcaacttgaa gaccacatac 2041 agatccaaga aacccgctaa gaacctcaag atgcccggcg tctactatgt ggacagaaga 2101 ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg agcagcacga ggtggctgtg 2161 gccagatact gcgacctccc tagcaaactg gggcacagat gagcggccgc gactctagat 2221 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct 2281 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc 2341 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 2401 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa 2461 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc 2521 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga 2581 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag 2641 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt 2701 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta 2761 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag 2821 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg 2881 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa 2941 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 3001 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct 3061 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 3121 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc 3181 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac 3241 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 3301 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 3361 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag 3421 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 3481 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 3541 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 3601 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 3661 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 3721 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 3781 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 3841 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 3901 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga 3961 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 4021 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 4081 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 4141 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 4201 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 4261 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 4321 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 4381 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga 4441 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt 4501 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc 4561 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg 4621 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta 4681 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 4741 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 4801 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 4861 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 4921 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 4981 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 5041 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 5101 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 5161 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 5221 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 5281 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 5341 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 5401 aggggggcgg agcctatgga aaaacgcc //