LOCUS Exported 6497 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Visualization of ER in mammalian cells using mGold . ACCESSION . VERSION . KEYWORDS pCMV-mGold-Calnexin-N-14 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6497) AUTHORS Lee J, Liu Z, Suzuki PH, Ahrens JF, Lai S, Lu X, Guan S, St-Pierre F TITLE Versatile phenotype-activated cell sorting. JOURNAL Sci Adv. 2020 Oct 23;6(43). pii: 6/43/eabb7438. doi: 10.1126/sciadv.abb7438. Print 2020 Oct. PUBMED 33097540 REFERENCE 2 (bases 1 to 6497) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6497 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 76..379 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 380..583 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 534..554 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 2452..2461 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2458..3177 /codon_start=1 /product="photostable derivative of mVenus (Lee et al., 2020)" /label=mGold /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2503..2524) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2764..2783) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3111..3132 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3300..3421 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3337..3356) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3391..3410 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3428..3883) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3565..3586) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3777..3796 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3910..4014 /gene="bla" /label=AmpR promoter promoter 4016..4373 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4039..4059) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4224..4359 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4286..4305 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4408..5202 /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(4462..4481) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5072..5091 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5390..5409) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5434..5481 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5810..6398 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6299..6318 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc 61 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 361 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca 601 gatccgctag cgctaccgga ctcagatctc gagaccgcca tggaagggaa gtggttgctg 661 tgtatgttac tggtgcttgg aactgctatt gttgaggctc atgatggaca tgatgatgat 721 gtgattgata ttgaggatga ccttgacgat gtcattgaag aggtagaaga ctcaaaacca 781 gataccactg ctcctccttc atctcccaag gttacttaca aagctccagt tccaacaggg 841 gaagtatatt ttgctgattc ttttgacaga ggaactctgt cagggtggat tttatccaaa 901 gccaagaaag acgataccga tgatgaaatt gccaaatatg atggaaagtg ggaggtagag 961 gaaatgaagg agtcaaagct tccaggtgat aaaggacttg tgttgatgtc tcgggccaag 1021 catcatgcca tctctgctaa actgaacaag cccttcctgt ttgacaccaa gcctctcatt 1081 gttcagtatg aggttaattt ccaaaatgga atagaatgtg gtggtgccta tgtgaaactg 1141 ctttctaaaa caccagaact caacctggat cagttccatg acaagacccc ttatacgatt 1201 atgtttggtc cagataaatg tggagaggac tataaactgc acttcatctt ccgacacaaa 1261 aaccccaaaa cgggtatcta tgaagaaaaa catgctaaga ggccagatgc agatctgaag 1321 acctatttta ctgataagaa aacacatctt tacacactaa tcttgaatcc agataatagt 1381 tttgaaatac tggttgacca atctgtggtg aatagtggaa atctgctcaa tgacatgact 1441 cctcctgtaa atccttcacg tgaaattgag gacccagaag accggaagcc cgaggattgg 1501 gatgaaagac caaaaatccc agatccagaa gctgtcaagc cagatgactg ggatgaagat 1561 gcccctgcta agattccaga tgaagaggcc acaaaacccg aaggctggtt agatgatgag 1621 cctgagtacg tacctgatcc agacgcagag aaacctgagg attgggatga agacatggat 1681 ggagaatggg aggctcctca gattgccaac cctagatgtg agtcagctcc tggatgtggt 1741 gtctggcagc gacctgtgat tgacaacccc aattataaag gcaaatggaa gcctcctatg 1801 attgacaatc ccagttacca gggaatctgg aaacccagga aaataccaaa tccagatttc 1861 tttgaagatc tggaaccttt cagaatgact ccttttagtg ctattggttt ggagctgtgg 1921 tccatgacct ctgacatttt ttttgacaac tttatcattt gtgctgatcg aagaatagtt 1981 gatgattggg ccaatgatgg atggggcctg aagaaagctg ctgatggggc tgctgagcca 2041 ggcgttgtgg ggcagatgaa cgaggcagct gaagagcgcc cgtggctgtg ggtagtctat 2101 attctaactg tagcccttcc tgtgttcctg gttatcctct tctgctgttc tggaaagaaa 2161 cagaccagtg gtatggagta taagaaaact gatgcacctc aaccggatgt gaaggaagag 2221 gaagaagaga aggaagagga aaaggacaag ggagatgagg aggaggaagg agaagagaaa 2281 cttgaagaga aacagaaaag tgatgctgaa gaagatggtg gcactgtcag tcaagaggag 2341 gaagacagaa aacctaaagc agaggaggat gaaattttga acagatcacc aagaaacaga 2401 aagccacgaa gagagggggg tgcgtcaggc ggcagcggat ctggaccggt cgccaccatg 2461 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 2521 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 2581 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 2641 gtgaccagcc tgggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag 2701 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 2761 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 2821 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 2881 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc 2941 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac 3001 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 3061 ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg 3121 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc 3181 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa 3241 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta 3301 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 3361 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 3421 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc 3481 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag 3541 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg 3601 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca 3661 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa 3721 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg 3781 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta 3841 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg 3901 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 3961 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag 4021 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc 4081 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt 4141 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 4201 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 4261 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 4321 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat 4381 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct 4441 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc 4501 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc 4561 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc 4621 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg 4681 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag 4741 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc 4801 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt 4861 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc 4921 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc 4981 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg 5041 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag 5101 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg 5161 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg 5221 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct 5281 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc 5341 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga 5401 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc 5461 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg 5521 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc 5581 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg 5641 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta 5701 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 5761 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 5821 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 5881 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 5941 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 6001 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 6061 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 6121 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 6181 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 6241 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 6301 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 6361 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 6421 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 6481 attctgtgga taaccgt //