LOCUS Exported 4793 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION In vivo visualization of actin cytoskeleton (can be used for colocalization studies). ACCESSION . VERSION . KEYWORDS ITPKA-mScarlet-I SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4793) AUTHORS Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J TITLE Robust and Bright Genetically Encoded Fluorescent Markers for Highlighting Structures and Compartments in Mammalian Cells JOURNAL bioRxiv 160374 REFERENCE 2 (bases 1 to 4793) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4793 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 191..211 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter complement(236..340) /gene="bla" /label=AmpR promoter rep_origin 367..822 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(454..473) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 664..685 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 829..950 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(840..859) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 894..913 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(1076..1771) /codon_start=1 /product="bright monomeric red fluorescent protein evolved from a synthetic template (Bindels et al., 2016)" /label=mScarlet-I /note="matures faster than mScarlet" /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH STGGMDELYK" primer_bind complement(1351..1370) /label=mCherry-F /note="mCherry, forward primer" primer_bind 1620..1638 /label=mCherry-R /note="mCherry, reverse primer" regulatory 1768..1777 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(1963..2166) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(1992..2012) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 2167..2470 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" rep_origin complement(2645..3233) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2725..2744) /label=pBR322ori-F /note="pBR322 origin, forward primer" polyA_signal 3562..3609 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" primer_bind 3634..3653 /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" CDS complement(3841..4635) /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(3952..3971) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 4562..4581 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(4670..234) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4684..26 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(4738..4757) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" ORIGIN 1 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac 61 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac 121 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg 181 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcaggactct 241 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 301 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 361 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 421 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 481 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 541 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 601 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 661 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg 721 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa 781 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttacgcctta agatacattg 841 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt 901 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca 961 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt 1021 aaaacctcta caaatgtggt atggctgatt atgatctaga gtcgcggccg ctttacttgt 1081 acagctcgtc catgccgccg gtggagtggc ggccctcgga gcgttcgtac tgttccacca 1141 cggtgtagtc ctcgttgtgg gaggtgatgt ccaacttgcg gtcgacgttg taggcgccgg 1201 gcatctgcac gggcttcttg gccttgtagg tggtcttgaa gtccgccagg tagcggccgc 1261 cgtccttcag gcgcagggcc atcttaatgt cgcccttcag cacgccgtcc tcggggtaca 1321 accgctcggt ggacgcttcc cagcccattg tcttcttctg cattacgggg ccgtcaggag 1381 ggaagttggt gccgcggagc ttcaccttgt agatcagggt gccgtcctcc agggaggtgt 1441 cctgggtcac ggtcacggcg ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc 1501 cctcggggaa ggactgctta tagtagtcgg ggatgtcggc ggggtgcttg atgaaggccc 1561 tggagccgta catgaactga ggggacagga tgtcccagga gaagggcagg gggccaccct 1621 tggtcacctt cagcttggcg gtctgggtgc cctcgtaggg gcggccctcg ccctcgccct 1681 cgatctcgaa ctcgtggccg ttcatggagc cctccatgtg caccttgaac cgcatgaact 1741 ccttgatcac tgcctcgccc ttgctcacca tggtggcgac cggtggatcc gatccagatc 1801 cgcccgcttc gaagagcagg cgcagctccc cgacgctccg gcgcggagcc cgctccaacc 1861 cggggctgca gggccccgcg ccccgtggtc gcgccatgcc cataagcttg agctcgagat 1921 ctgagtccgg tagcgctagc ggatctgacg gttcactaaa ccagctctgc ttatatagac 1981 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt 2041 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag 2101 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg 2161 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca 2221 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg 2281 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata 2341 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat 2401 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag 2461 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta 2521 ttaataacta atgcatggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 2581 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 2641 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 2701 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 2761 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 2821 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 2881 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 2941 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 3001 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 3061 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 3121 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 3181 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 3241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 3301 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 3361 ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga 3421 cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag 3481 aaacgcgggc gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct 3541 gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt 3601 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc 3661 tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc 3721 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga 3781 aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc 3841 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat 3901 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg 3961 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa 4021 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac 4081 gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc 4141 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt 4201 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag 4261 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg 4321 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc 4381 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg 4441 cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac 4501 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg 4561 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc 4621 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcgatct ttgcaaaagc 4681 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct 4741 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcg //