LOCUS Exported 4790 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Visualization of the Endoplasmic Reticulum. ACCESSION . VERSION . KEYWORDS ER-mScarletI SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4790) 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 4790) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4790 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 115..418 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 419..622 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 573..593 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" sig_peptide 695..748 /product="signal sequence of chicken lysozyme" /label=lysozyme signal sequence regulatory 797..806 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 803..1498 /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(936..954) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1204..1223 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 1632..1753 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1669..1688) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1723..1742 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1760..2215) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1897..1918) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2109..2128 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2242..2346 /gene="bla" /label=AmpR promoter promoter 2348..2705 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2371..2391) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2556..2691 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2618..2637 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2740..3534 /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(2794..2813) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3404..3423 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3722..3741) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3766..3813 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4142..4730 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4631..4650 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta 61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac 121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc 181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt 241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac 301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac 361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt 421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc 481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt 541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg 601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac 661 tcagatctcg agctcaagct tcgaattcgc caccatgagg tctttgctaa tcttggtgct 721 ttgcttcctg cccctggctg ctctggggaa agtctttgga cgatgtgagc tggcagcggc 781 tatgaagcca ccggtcgcca ccatggtgag caagggcgag gcagtgatca aggagttcat 841 gcggttcaag gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga 901 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg 961 ccccctgccc ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt 1021 catcaagcac cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa 1081 gtgggagcgc gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc 1141 cctggaggac ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga 1201 cggccccgta atgcagaaga agacaatggg ctgggaagca tccaccgagc ggttgtaccc 1261 cgaggacggc gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggccg 1321 ctacctggcg gacttcaaga ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc 1381 ctacaacgtc gaccgcaagt tggacatcac ctcccacaac gaggactaca ccgtggtgga 1441 acagtacgaa cgctccgagg gccgccactc caccggcggc atggacgagc tgtacaagga 1501 tgaattgtaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt 1561 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa 1621 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 1681 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 1741 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat 1801 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa 1861 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta 1921 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca 1981 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat 2041 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg 2101 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc 2161 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt 2221 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 2281 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 2341 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt 2401 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 2461 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 2521 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 2581 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 2641 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 2701 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 2761 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 2821 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 2881 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta 2941 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 3001 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 3061 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 3121 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 3181 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 3241 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc 3301 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 3361 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 3421 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 3481 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 3541 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt 3601 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga 3661 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa 3721 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac 3781 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct 3841 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct 3901 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg 3961 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc 4021 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc 4081 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 4141 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 4201 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 4261 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 4321 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 4381 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 4441 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 4501 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 4561 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 4621 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 4681 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 4741 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg //