LOCUS Exported 6024 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION In vivo visualization of microtubules (can be used for colocalization studies). ACCESSION . VERSION . KEYWORDS pmScarlet-i_alphaTubulin_C1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6024) 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 6024) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6024 /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 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1308 /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(746..764) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1014..1033 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2812..2933 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2849..2868) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2903..2922 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2940..3395) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3077..3098) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3289..3308 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3422..3526 /gene="bla" /label=AmpR promoter promoter 3528..3885 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3551..3571) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3736..3871 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3798..3817 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3920..4714 /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(3974..3993) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4584..4603 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4902..4921) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4946..4993 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5322..5910 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5811..5830 /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 ccggtcgcca ccatggtgag caagggcgag gcagtgatca aggagttcat gcggttcaag 661 gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc 721 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc 781 ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt catcaagcac 841 cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc 901 gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac 961 ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta 1021 atgcagaaga agacaatggg ctgggaagca tccaccgagc ggttgtaccc cgaggacggc 1081 gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg 1141 gacttcaaga ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc 1201 gaccgcaagt tggacatcac ctcccacaac gaggactaca ccgtggtgga acagtacgaa 1261 cgctccgagg gccgccactc caccggcggc atggacgagc tgtacaagtc cggactcaga 1321 tctcgagtgc gtgagtgcat ctccatccac gttggccagg ctggtgtcca gattggcaat 1381 gcctgctggg agctctactg cctggaacac ggcatccagc ccgatggcca gatgccaagt 1441 gacaagacca ttgggggagg agatgactcc ttcaacacct tcttcagtga gacgggcgct 1501 ggcaagcacg tgccccgggc tgtgtttgta gacttggaac ccacagtcat tgatgaagtt 1561 cgcactggca cctaccgcca gctcttccac cctgagcagc tcatcacagg caaggaagat 1621 gctgccaata actatgcccg agggcactac accattggca aggagatcat tgaccttgtg 1681 ttggaccgaa ttcgcaagct ggctgaccag tgcaccggtc ttcagggctt cttggttttc 1741 cacagctttg gtgggggaac tggttctggg ttcacctccc tgctcatgga acgtctctca 1801 gttgattatg gcaagaagtc caagctggag ttctccattt acccagcacc ccaggtttcc 1861 acagctgtag ttgagcccta caactccatc ctcaccaccc acaccaccct ggagcactct 1921 gattgtgcct tcatggtaga caatgaggcc atctatgaca tctgtcgtag aaacctcgat 1981 atcgagcgcc caacctacac taaccttaac cgccttatta gccagattgt gtcctccatc 2041 actgcttccc tgagatttga tggagccctg aatgttgacc tgacagaatt ccagaccaac 2101 ctggtgccct acccccgcat ccacttccct ctggccacat atgcccctgt catctctgct 2161 gagaaagcct accatgaaca gctttctgta gcagagatca ccaatgcttg ctttgagcca 2221 gccaaccaga tggtgaaatg tgaccctcgc catggtaaat acatggcttg ctgcctgttg 2281 taccgtggtg acgtggttcc caaagatgtc aatgctgcca ttgccaccat caaaaccaag 2341 cgcagcatcc agtttgtgga ttggtgcccc actggcttca aggttggcat caactaccag 2401 cctcccactg tggtgcctgg tggagacctg gccaaggtac agagagctgt gtgcatgctg 2461 agcaacacca cagccattgc tgaggcctgg gctcgcctgg accacaagtt tgacctgatg 2521 tatgccaagc gtgcctttgt tcactggtac gtgggtgagg ggatggagga aggcgagttt 2581 tcagaggccc gtgaagatat ggctgccctt gagaaggatt atgaggaggt tggtgtggat 2641 tctgttgaag gagagggtga ggaagaagga gaggaatact aaggatccac cggatctaga 2701 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc 2761 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt 2821 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 2881 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta 2941 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt 3001 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat 3061 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa 3121 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta 3181 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc 3241 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc 3301 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac 3361 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt 3421 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag 3481 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag 3541 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 3601 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 3661 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 3721 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 3781 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc 3841 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac 3901 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc 3961 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 4021 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 4081 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg 4141 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 4201 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 4261 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 4321 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 4381 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 4441 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc 4501 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 4561 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg 4621 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 4681 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc 4741 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 4801 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 4861 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca 4921 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt 4981 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca 5041 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca 5101 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc 5161 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 5221 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 5281 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 5341 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 5401 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 5461 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 5521 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 5581 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 5641 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 5701 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 5761 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 5821 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 5881 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 5941 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 6001 gataaccgta ttaccgccat gcat //