LOCUS Exported 5007 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expression of fusion incompetent VAMP3 (del71) C-terminally conjugated to mCherry.. ACCESSION . VERSION . KEYWORDS VAMP3(Delta-71)-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5007) AUTHORS Verboogen DRJ, Gonzalez Mancha N, Ter Beest M, van den Bogaart G TITLE Fluorescence lifetime imaging microscopy reveals rerouting of SNARE trafficking driving dendritic cell activation. JOURNAL Elife. 2017 May 19;6. pii: e23525. doi: 10.7554/eLife.23525. PUBMED 28524818 REFERENCE 2 (bases 1 to 5007) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5007 /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 956..965 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 962..1672 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1107..1125) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1375..1394 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 1795..1916 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1832..1851) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1886..1905 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1923..2378) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2060..2081) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2272..2291 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2405..2509 /gene="bla" /label=AmpR promoter promoter 2511..2868 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2534..2554) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2719..2854 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2781..2800 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2903..3697 /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(2957..2976) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3567..3586 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3885..3904) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3929..3976 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4305..4893 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4794..4813 /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 atctcgagct caagcttcga attcatgtct acaggggtgc cttcagggtc 661 cagtgctgcc actggcagta atcgaagact ccagcagaca cagaatcaag tagatgaggt 721 ggtggacatc atgagagtca atgtggataa ggtgttagaa agagaccaga agctctcaga 781 gctagacgac cgcgcagatg cactgcaggc aggcgcctcg cagtttgaaa caagtgctgc 841 caagaagaga aagtattggt ggaagaactg caagatgtgg gcgataggga tcagtgtcct 901 ggtgattatt gtcattatca tcattgtgtg gtgtgtctct aaggatccac cggtcgccac 961 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa 1021 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg 1081 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc 1141 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca 1201 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg 1261 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga 1321 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt 1381 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg 1441 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc 1501 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt 1561 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga 1621 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aaagcggccg 1681 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac 1741 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg 1801 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 1861 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc 1921 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 1981 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 2041 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 2101 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 2161 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 2221 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 2281 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 2341 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa 2401 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 2461 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga 2521 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 2581 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 2641 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 2701 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 2761 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta 2821 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga 2881 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 2941 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 3001 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 3061 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac 3121 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 3181 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 3241 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 3301 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 3361 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 3421 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 3481 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 3541 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 3601 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 3661 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg 3721 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat 3781 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 3841 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag 3901 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt 3961 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc 4021 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc 4081 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata 4141 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg 4201 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 4261 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 4321 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 4381 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 4441 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 4501 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 4561 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 4621 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 4681 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 4741 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 4801 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 4861 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 4921 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct 4981 gtggataacc gtattaccgc catgcat //