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" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 754..763 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 760..1455 /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(893..911) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1161..1180 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 1581..1702 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1618..1637) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1672..1691 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1709..2164) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1846..1867) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2058..2077 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2191..2295 /gene="bla" /label=AmpR promoter promoter 2297..2654 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2320..2340) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2505..2640 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2567..2586 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2689..3483 /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(2743..2762) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3353..3372 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3671..3690) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3715..3762 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4091..4679 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4580..4599 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atctcgagct caagcttatg ggcatggcgc gaccacgggg cgcggggccc 661 tgcagccccg ggttggagcg ggctccgcgc cggagcgtcg gggagctgcg cctgctcttc 721 gaagcgggcg gatctggatc ggatccaccg gtcgccacca tggtgagcaa gggcgaggca 781 gtgatcaagg agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag 841 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg 901 aaggtgacca agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg 961 tacggctcca gggccttcat caagcacccc gccgacatcc ccgactacta taagcagtcc 1021 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc 1081 gtgacccagg acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc 1141 accaacttcc ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcatcc 1201 accgagcggt tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc 1261 ctgaaggacg gcggccgcta cctggcggac ttcaagacca cctacaaggc caagaagccc 1321 gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag 1381 gactacaccg tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcatg 1441 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg 1501 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa 1561 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca 1621 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 1681 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc 1741 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc 1801 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag 1861 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc 1921 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa 1981 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg 2041 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt 2101 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc 2161 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 2221 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 2281 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt 2341 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 2401 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 2461 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 2521 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 2581 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 2641 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa 2701 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg 2761 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc 2821 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca 2881 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc 2941 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca 3001 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat 3061 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca 3121 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg 3181 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc 3241 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct 3301 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct 3361 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac 3421 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc 3481 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag 3541 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 3601 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg 3661 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa 3721 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt 3781 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg 3841 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca 3901 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt 3961 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga 4021 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca 4081 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 4141 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 4201 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag 4261 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 4321 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 4381 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 4441 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 4501 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 4561 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 4621 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 4681 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 4741 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat //