LOCUS Exported 4849 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION In vivo visualization of the ER (can be used for colocalization studies and the OSER assay). ACCESSION . VERSION . KEYWORDS CytERM-mTurquoise2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4849) TITLE Gadella lab plasmids REFERENCE 2 (bases 1 to 4849) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4849 /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 790..799 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 796..1515 /codon_start=1 /product="enhanced monomeric variant of CFP (Goedhart et al., 2012)" /label=mTurquoise2 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(841..862) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1102..1121) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1449..1470 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1637..1758 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1674..1693) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1728..1747 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1765..2220) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1902..1923) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2114..2133 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2247..2351 /gene="bla" /label=AmpR promoter promoter 2353..2710 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2376..2396) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2561..2696 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2623..2642 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2745..3539 /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(2799..2818) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3409..3428 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3727..3746) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3771..3818 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4147..4735 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4636..4655 /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 atctggtacc ccttgcagct tccagttgtc aggaagagaa ggctcccatg 661 gatcctgtgg tggtgctggg gctctgtctc tcctgtttgc ttctcctttc actctggaaa 721 cagagctatg ggggagggaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat 781 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc 841 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag 901 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc 961 gtgccctggc ccaccctcgt gaccaccctg tcctggggcg tgcagtgctt cgcccgctac 1021 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag 1081 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc 1141 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc 1201 aacatcctgg ggcacaagct ggagtacaac tactttagcg acaacgtcta tatcaccgcc 1261 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc 1321 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg 1381 ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag 1441 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac 1501 gagctgtaca agtaagcggc cgcgactcta gatcataatc agccatacca catttgtaga 1561 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa 1621 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag 1681 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 1741 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt 1801 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt 1861 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc 1921 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg 1981 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac 2041 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg 2101 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag 2161 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt 2221 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 2281 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 2341 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg 2401 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 2461 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 2521 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 2581 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 2641 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 2701 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 2761 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 2821 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 2881 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc 2941 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 3001 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 3061 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 3121 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 3181 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 3241 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg 3301 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 3361 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 3421 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 3481 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 3541 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 3601 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 3661 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag 3721 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa 3781 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca 3841 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc 3901 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt 3961 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa 4021 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa 4081 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg 4141 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc 4201 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac 4261 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca 4321 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt 4381 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc 4441 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg 4501 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc 4561 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac 4621 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct 4681 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc 4741 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt 4801 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat //