LOCUS Exported 5061 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION In vivo visualization of the Golgi apparatus (can be used for colocalization studies). ACCESSION . VERSION . KEYWORDS pmScarlet-i_Giantin_C1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5061) AUTHORS Bindels DS, Haarbosch L, van Weeren L, Postma M, Wiese KE, Mastop M, Aumonier S, Gotthard G, Royant A, Hink MA, Gadella TW Jr TITLE mScarlet: a bright monomeric red fluorescent protein for cellular imaging. JOURNAL Nat Methods. 2016 Nov 21. doi: 10.1038/nmeth.4074. PUBMED 27869816 REFERENCE 2 (bases 1 to 5061) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5061 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 59..362 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 363..566 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 517..537 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 605..614 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 611..1306 /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(744..762) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1012..1031 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 1847..1968 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1884..1903) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1938..1957 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1975..2430) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2112..2133) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2324..2343 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2457..2561 /gene="bla" /label=AmpR promoter promoter 2563..2920 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2586..2606) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2771..2906 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2833..2852 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2955..3749 /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(3009..3028) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3619..3638 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3937..3956) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3981..4028 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4357..4945 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4846..4865 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc 601 ggtcgccacc atggtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt 661 gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg 721 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt 781 ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca tcaagcaccc 841 cgccgacatc cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt 901 gatgaacttc gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg 961 caccctgatc tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat 1021 gcagaagaag acaatgggct gggaagcatc caccgagcgg ttgtaccccg aggacggcgt 1081 gctgaagggc gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga 1141 cttcaagacc acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga 1201 ccgcaagttg gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg 1261 ctccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtccg gactcagatc 1321 tcgaggagaa ccgcagcaaa gcttttctga agctcagcag cagctatgca acaccagaca 1381 ggaagtgaat gaattaagga agctgctgga agaagaacga gaccaaagag tggctgctga 1441 gaatgctctc tctgtggccg aggagcagat cagacggtta gagcacagtg aatgggactc 1501 ttcccggact cctatcattg gctcctgtgg cactcaggag caggcactgt taatagatct 1561 tacaagcaac agttgtcgaa ggacccggag tggcgttgga tggaagcgag tcctgcgttc 1621 actctgtcat tcacggaccc gagtgccact tctagcagcc atctactttc taatgattca 1681 tgtcctgctc attctgtgtt ttacgggcca tctataggga tccaccggat ctagataact 1741 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca 1801 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc 1861 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 1921 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg 1981 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta 2041 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt 2101 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca 2161 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa 2221 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat 2281 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag 2341 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg 2401 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg 2461 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 2521 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca 2581 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag 2641 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc 2701 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct 2761 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 2821 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa 2881 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat 2941 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 3001 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 3061 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 3121 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 3181 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 3241 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 3301 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 3361 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 3421 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 3481 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 3541 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 3601 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 3661 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 3721 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca 3781 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 3841 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 3901 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc 3961 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc 4021 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag 4081 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc 4141 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg 4201 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 4261 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 4321 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 4381 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 4441 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 4501 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 4561 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 4621 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 4681 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 4741 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 4801 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 4861 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 4921 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 4981 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa 5041 ccgtattacc gccatgcatt a //