LOCUS       sequence-451926-        5344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes mutant siRNA resistant SigmaR1 protein (transmembrane region
            deletion + substitutions) labeled with mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5344)
  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 5344)
  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 ss-SigmaR1DTM-mNeonGreen R119A...
FEATURES             Location/Qualifiers
     source          1..5344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      262..271
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      523..532
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1246..1449)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1275..1295)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1450..1753
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1928..2516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2008..2027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2845..2892
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2917..2936
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3124..3918)
                     /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(3124..3918)
                     /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(3235..3254)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3845..3864
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3953..4310)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3967..4102
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4119..4310
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4267..4287
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4312..4416)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4443..4898
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4530..4549)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4740..4761
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4905..5026
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4916..4935)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4970..4989
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5163..5344,1..526))
                     /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 gcttgccatt tccagtggtg taactccact taaaggtact gatgatggtt ttgtcgttgg
       61 ggtaagtctt cttcgacctg caccagtccg cagcggtcag cgagttggtc atcacaggac
      121 cgtcagcagg gaaaccagtc cccttcacct gggcctctcc tttgatgtgg cttccctcgt
      181 aggtgtagcg gtagttaaca gtaagggagg caccatcttc aaactgcatt gtgcgatgga
      241 cttggtagcc ggagccatct accatggcgg cctggaaagg cgacatcccg tcagggtagg
      301 gcaggtactg atggaagcca tacccgatat gagggaccag aatccagggg gagaactgga
      361 ggtcaccctt ggtggacttc aggtttaact cctcataacc atcatttgga ttgccggtgc
      421 cctgacccac catgtcaaag tccacaccgt tgatggagcc aaagatgtgt aactcatgtg
      481 tcgctgggag agaggccatg ttatcctcct cgcccttgct caccatggtg gcgaccggtg
      541 gatcccgggc ccgcggtacc ctagggtcct ggccaaagag gtatgtcgtc aattccaagc
      601 ggaggccccg agcataggag cgaagagtat agaagagggt gaggaagtcc tgggtgctag
      661 cgacagtgtc ggccagcgcg aaggccaggg tggatgggat gacgccccgg ccgtactcga
      721 ccatccaagt attgggcccc cactccacag ctgttgcctc accaggcccg tgtactaccg
      781 tctcccctgg gtagaagacc tcacttttgg tggtgccctc tctccactgg tggaaggtgc
      841 cagagatgat ggtatccgag atctcagccc agtaagcccc cgagtggccg cgggagccca
      901 aggcggtgcc gaagagcagc acatactcgg acagcgaggc gtgcagaagg cacatggcgc
      961 ccatccatcc accagcgttg acgaacaccc actgcagctc ctcgtcgggc agcacgtggc
     1021 ctgggtgcag ccgccgcagc tccacgatca gtctgctaaa tgcgagttcg tggtccagcc
     1081 cagcgtactg ccgcgccaac tgcgctatct cttcgcgctg gaagacgaag ctctgcgtac
     1141 cggcccgcgc cgcgctgagc agcagcagca tcgcggccac cagggagagc ttcatctcga
     1201 gatctgagtc cggtagcgct agcggatctg acggttcact aaaccagctc tgcttatata
     1261 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     1321 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     1381 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     1441 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     1501 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     1561 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     1621 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     1681 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     1741 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     1801 ctattaataa ctaatgcatg gcggtaatac ggttatccac agaatcaggg gataacgcag
     1861 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1921 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1981 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2041 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2101 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2161 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2221 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2281 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2341 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2401 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2461 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2521 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2581 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2641 gttttaaatc aatctaaagt atatatgagt aacctgaggc tatggcaggg cctgccgccc
     2701 cgacgttggc tgcgagccct gggccttcac ccgaacttgg ggggtggggt ggggaaaagg
     2761 aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc
     2821 cctgggaccg aaccccgcgt ttatgaacaa acgacccaac accgtgcgtt ttattctgtc
     2881 tttttattgc cgtcatagcg cgggttcctt ccggtattgt ctccttccgt gtttcagtta
     2941 gcctccccct agggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     3001 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     3061 cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc
     3121 cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     3181 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     3241 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
     3301 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
     3361 cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg
     3421 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     3481 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     3541 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     3601 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     3661 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     3721 ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     3781 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     3841 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     3901 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa
     3961 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
     4021 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     4081 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     4141 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     4201 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     4261 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag gactcttcct
     4321 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4381 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4441 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4501 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4561 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4621 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4681 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4741 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4801 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4861 taacgcgaat tttaacaaaa tattaacgct tacaatttac gccttaagat acattgatga
     4921 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     4981 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     5041 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     5101 cctctacaaa tgtggtatgg ctgattatga tctagagtcg cggccgctta tagctcgtct
     5161 ttcttgtaca gctcgtccat gcccatcaca tcggtaaagg ccttttgcca ctccttgaag
     5221 ttgagctcgg tcttggagtg cttgagctcc gtcttacgga acacgtacat cggctggttc
     5281 ttcagatagt tagccgccat tggcttggca aaggtgtagg tggtccgcgc agtgctccgg
     5341 tagc
//
