LOCUS Exported 4743 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION In vivo visualization of the plasma membrane (can be used for colocalization studies). ACCESSION . VERSION . KEYWORDS Lck-mScarlet-I SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4743) 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 4743) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4743 /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 595..604 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 640..649 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 646..1341 /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(779..797) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1047..1066 /label=mCherry-F /note="mCherry, forward primer" misc_feature 1342..1407 /label=MCS /note="multiple cloning site" polyA_signal 1531..1652 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1568..1587) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1622..1641 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1659..2114) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1796..1817) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2008..2027 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2141..2245 /gene="bla" /label=AmpR promoter promoter 2247..2604 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2270..2290) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2455..2590 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2517..2536 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2639..3433 /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(2693..2712) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3303..3322 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3621..3640) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3665..3712 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4041..4629 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4530..4549 /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 gctagccacc 601 atgggctgcg tgtgcagcag caaccccgag ctaccggtcg ccaccatggt gagcaagggc 661 gaggcagtga tcaaggagtt catgcggttc aaggtgcaca tggagggctc catgaacggc 721 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc 781 aagctgaagg tgaccaaggg tggccccctg cccttctcct gggacatcct gtcccctcag 841 ttcatgtacg gctccagggc cttcatcaag caccccgccg acatccccga ctactataag 901 cagtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgcc 961 gtgaccgtga cccaggacac ctccctggag gacggcaccc tgatctacaa ggtgaagctc 1021 cgcggcacca acttccctcc tgacggcccc gtaatgcaga agaagacaat gggctgggaa 1081 gcatccaccg agcggttgta ccccgaggac ggcgtgctga agggcgacat taagatggcc 1141 ctgcgcctga aggacggcgg ccgctacctg gcggacttca agaccaccta caaggccaag 1201 aagcccgtgc agatgcccgg cgcctacaac gtcgaccgca agttggacat cacctcccac 1261 aacgaggact acaccgtggt ggaacagtac gaacgctccg agggccgcca ctccaccggc 1321 ggcatggacg agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca 1381 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat 1441 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg 1501 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac 1561 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 1621 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt 1681 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat 1741 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt 1801 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt 1861 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag 1921 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg 1981 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc 2041 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc 2101 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 2161 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 2221 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt 2281 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 2341 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg 2401 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg 2461 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 2521 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt 2581 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat 2641 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 2701 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 2761 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 2821 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 2881 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 2941 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 3001 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 3061 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 3121 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg 3181 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 3241 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 3301 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 3361 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 3421 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg 3481 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt 3541 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc 3601 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct 3661 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc 3721 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa 3781 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg 3841 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt 3901 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat 3961 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 4021 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 4081 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 4141 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag 4201 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 4261 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 4321 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 4381 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 4441 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 4501 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 4561 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 4621 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 4681 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg 4741 cat //