LOCUS       sequence-451927-        5368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes mutant siRNA resistant SigmaR1 protein (substitutions) 
            labeled with mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5368)
  AUTHORS   Sawyer EM, Jensen LE, Meehl JB, Larsen KP, Petito DA, Hurley JH, 
            Voeltz GK
  TITLE     SigmaR1 shapes rough endoplasmic reticulum membrane sheets.
  JOURNAL   Dev Cell. 2024 Jul 2:S1534-5807(24)00382-4. doi: 
            10.1016/j.devcel.2024.06.005.
  PUBMED    38971154
REFERENCE   2  (bases 1 to 5368)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dev Cell. 
            2024 Jul 2:S1534-5807(24)00382-4. doi: 
            10.1016/j.devcel.2024.06.005."
COMMENT     Sequence Label: siRNA-resistant SigmaR1-mNeonGreen R119A F191A
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      238..247
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      499..508
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1258..1461)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        1462..1765
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1940..2528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2020..2039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2857..2904
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2929..2948
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3136..3930)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3136..3930)
                     /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(3247..3266)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3857..3876
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3965..4322)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3979..4114
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4131..4322
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4279..4299
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4324..4428)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4455..4910
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4542..4561)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4752..4773
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4917..5038
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4928..4947)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4982..5001
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5160..5368,1..502))
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
ORIGIN
        1 tccacttaaa ggtactgatg atggttttgt cgttggggta agtcttcttc gacctgcacc
       61 agtccgcagc ggtcagcgag ttggtcatca caggaccgtc agcagggaaa ccagtcccct
      121 tcacctgggc ctctcctttg atgtggcttc cctcgtaggt gtagcggtag ttaacagtaa
      181 gggaggcacc atcttcaaac tgcattgtgc gatggacttg gtagccggag ccatctacca
      241 tggcggcctg gaaaggcgac atcccgtcag ggtagggcag gtactgatgg aagccatacc
      301 cgatatgagg gaccagaatc cagggggaga actggaggtc acccttggtg gacttcaggt
      361 ttaactcctc ataaccatca tttggattgc cggtgccctg acccaccatg tcaaagtcca
      421 caccgttgat ggagccaaag atgtgtaact catgtgtcgc tgggagagag gccatgttat
      481 cctcctcgcc cttgctcacc atggtggcga ccggtggatc ccgggcccgc ggtaccctag
      541 ggtcctggcc aaagaggtat gtcgtcaatt ccaagcggag gccccgagca taggagcgaa
      601 gagtatagaa gagggtgagg aagtcctggg tgctagcgac agtgtcggcc agcgcgaagg
      661 ccagggtgga tgggatgacg ccccggccgt actcgaccat ccaagtattg ggcccccact
      721 ccacagctgt tgcctcacca ggcccgtgta ctaccgtctc ccctgggtag aagacctcac
      781 ttttggtggt gccctctctc cactggtgga aggtgccaga gatgatggta tccgagatct
      841 cagcccagta agcccccgag tggccgcggg agcccaaggc ggtgccgaag agcagcacat
      901 actcggacag cgaggcgtgc agaaggcaca tggcgcccat ccatccacca gcgttgacga
      961 acacccactg cagctcctcg tcgggcagca cgtggcctgg gtgcagccgc cgcagctcca
     1021 cgatcagtct gctaaatgcg agttcgtggt ccagcccagc gtactgccgc gccaactgcg
     1081 ctatctcttc gcgctggaag acgaagctct gcgtacccag ccagagccag acgacctggg
     1141 tcagcaccgc tgcgacagcc aggagcagcg cggcccacgc ccaccgccgg cccacggccc
     1201 actgcatctc gagatctgag tccggtagcg ctagcggatc tgacggttca ctaaaccagc
     1261 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     1321 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     1381 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     1441 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     1501 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     1561 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     1621 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     1681 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     1741 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     1801 cgtaattgat tactattaat aactaatgca tggcggtaat acggttatcc acagaatcag
     1861 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1921 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1981 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2041 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2101 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2161 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2221 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2281 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2341 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2401 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2461 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2521 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2581 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2641 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaacctgag gctatggcag
     2701 ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc acccgaactt ggggggtggg
     2761 gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga
     2821 cagagtgcca gccctgggac cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg
     2881 ttttattctg tctttttatt gccgtcatag cgcgggttcc ttccggtatt gtctccttcc
     2941 gtgtttcagt tagcctcccc ctagggtggg cgaagaactc cagcatgaga tccccgcgct
     3001 ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg
     3061 cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga
     3121 accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
     3181 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
     3241 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
     3301 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
     3361 atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa
     3421 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
     3481 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
     3541 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
     3601 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
     3661 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
     3721 cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt
     3781 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
     3841 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
     3901 acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc
     3961 gatctttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag
     4021 aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag
     4081 aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg
     4141 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     4201 ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     4261 agcctgggga ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc
     4321 aggactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4381 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4441 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4501 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4561 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4621 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4681 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4741 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4801 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4861 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt acgccttaag
     4921 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4981 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     5041 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     5101 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatctagagt cgcggccgct
     5161 tacttgtaca gctcgtccat gcccatcaca tcggtaaagg ccttttgcca ctccttgaag
     5221 ttgagctcgg tcttggagtg cttgagctcc gtcttacgga acacgtacat cggctggttc
     5281 ttcagatagt tagccgccat tggcttggca aaggtgtagg tggtccgcgc agtgctccgg
     5341 tagcgcttgc catttccagt ggtgtaac
//
