LOCUS Exported 4839 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Fluorescent labeling of ER membrane in mammalian cells. ACCESSION . VERSION . KEYWORDS SNAP-Sec61B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4839) TITLE Fission and fusion machineries converge at ER contact sites to regulate mitochondrial morphology JOURNAL Unpublished REFERENCE 2 (bases 1 to 4839) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4839 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 547..556 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 553..1098 /codon_start=1 /product="human O6-alkylguanine-DNA-alkyltransferase variant that forms covalent bonds with benzylguanine derivatives" /label=SNAP-tag(R) /note="fast-reacting variant, optimized for mammalian expression" /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA VKEWLLAHEGHRLGKPGLG" CDS 1126..1416 /codon_start=1 /gene="SEC61B" /product="human Sec61-beta subunit of the translocon in the endoplasmic reticulum" /label=Sec61-beta /translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLMFIASVFMLHIWGKYTRS" polyA_signal 1567..1688 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1604..1623) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1658..1677 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1695..2150) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1832..1853) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2044..2063 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2177..2281 /gene="bla" /label=AmpR promoter promoter 2283..2640 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2306..2326) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2491..2626 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2553..2572 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2675..3469 /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(2729..2748) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3339..3358 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3657..3676) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3701..3748 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4077..4665 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4566..4585 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 541 ccggtcgcca ccatggacaa agactgcgaa atgaagcgca ccaccctgga tagccctctg 601 ggcaagctgg aactgtctgg gtgcgaacag ggcctgcacc gtatcatctt cctgggcaaa 661 ggaacatctg ccgccgacgc cgtggaagtg cctgccccag ccgccgtgct gggcggacca 721 gagccactga tgcaggccac cgcctggctc aacgcctact ttcaccagcc tgaggccatc 781 gaggagttcc ctgtgccagc cctgcaccac ccagtgttcc agcaggagag ctttacccgc 841 caggtgctgt ggaaactgct gaaagtggtg aagttcggag aggtcatcag ctacagccac 901 ctggccgccc tggccggcaa tcccgccgcc accgccgccg tgaaaaccgc cctgagcgga 961 aatcccgtgc ccattctgat cccctgccac cgggtggtgc agggcgacct ggacgtgggg 1021 ggctacgagg gcgggctcgc cgtgaaagag tggctgctgg cccacgaggg ccacagactg 1081 ggcaagcctg ggctgggtag cggtagcggt ggcagctctc gagctatgcc tggtccgacc 1141 cccagtggca ctaacgtggg atcctcaggg cgctctccca gcaaagcagt ggccgcccgg 1201 gcggcgggat ccactgtccg gcagaggaaa aatgccagct gtgggacaag gagtgcaggc 1261 cgcacaacct cggcaggcac cggggggatg tggcgattct acacagaaga ttcacctggg 1321 ctcaaagttg gccctgttcc agtattggtt atgagtctta tgttcatcgc ttctgtattt 1381 atgttgcaca tttggggcaa gtacactcgt tcgtagtcgt gaggtaccgc gggcccggga 1441 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt 1501 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt 1561 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 1621 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 1681 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg 1741 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa 1801 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa 1861 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg 1921 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa 1981 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa 2041 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct 2101 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac 2161 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 2221 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 2281 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 2341 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 2401 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 2461 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 2521 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 2581 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 2641 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 2701 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 2761 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 2821 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg 2881 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 2941 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 3001 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 3061 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 3121 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 3181 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg 3241 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 3301 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 3361 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 3421 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 3481 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 3541 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 3601 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa 3661 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat 3721 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac 3781 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc 3841 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca 3901 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt 3961 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 4021 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 4081 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 4141 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 4201 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 4261 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 4321 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 4381 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 4441 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 4501 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 4561 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 4621 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 4681 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 4741 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat 4801 tacggggtca ttagttcata gcccatatat ggagttccg //