LOCUS Exported 5023 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Visualization of Golgi in mammalian cells using mGold. ACCESSION . VERSION . KEYWORDS pCMV-mGold-MannII-N-10 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5023) 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 5023) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5023 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 68..371 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 372..575 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 526..546 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 970..979 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 976..1695 /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(1021..1042) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1282..1301) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1629..1650 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1818..1939 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1855..1874) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1909..1928 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1946..2401) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2083..2104) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2295..2314 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2428..2532 /gene="bla" /label=AmpR promoter promoter 2534..2891 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2557..2577) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2742..2877 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2804..2823 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2926..3720 /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(2980..2999) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3590..3609 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3908..3927) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3952..3999 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4328..4916 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4817..4836 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg 61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc 121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt 181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc 241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact 421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa 481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta 541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct 601 agcgccacca tgaagttaag tcgccagttc accgtgtttg gcagcgcgat cttctgcgtc 661 gtaatcttct cactctacct gatgctggac aggggtcact tggactaccc tcggggcccg 721 cgccaggagg gctcctttcc gcagggccag ctttcaatat tgcaagaaaa gattgaccat 781 ttggagcgtt tgctcgctga gaacaacgag atcatctcaa atatcagaga ctcagtcatc 841 aacctgagcg agtctgtgga ggacggcccg cgggggtcac caggcaacgc cagccaaggc 901 tccatccacc tccactcgcc acagttggcc ctgcaggctg accccagctc cggatcggat 961 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc 1021 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag 1081 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc 1141 gtgccctggc ccaccctcgt gaccagcctg ggctacggcc tgcagtgctt cgcccgctac 1201 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag 1261 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc 1321 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc 1381 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc 1441 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc 1501 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg 1561 ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag 1621 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac 1681 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag 1741 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga 1801 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata 1861 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 1921 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg 1981 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct 2041 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt 2101 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat 2161 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca 2221 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac 2281 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta 2341 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg 2401 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata 2461 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga 2521 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg 2581 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 2641 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 2701 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 2761 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 2821 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 2881 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat 2941 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca 3001 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg 3061 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg 3121 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact 3181 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct 3241 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg 3301 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt 3361 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc 3421 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc 3481 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga 3541 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc 3601 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt 3661 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga 3721 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 3781 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 3841 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga 3901 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa 3961 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc 4021 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt 4081 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg 4141 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa 4201 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca 4261 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 4321 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 4381 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 4441 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc 4501 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 4561 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 4621 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 4681 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 4741 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 4801 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 4861 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 4921 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 4981 ttcctgcgtt atcccctgat tctgtggata accgtattac cgc //