LOCUS       sequence-451909-        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 a5mut (F196...
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     565..584
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(883..902)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    927..974
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1303..1891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1792..1811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2066..2369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2370..2573
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3323..3332
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3329..4039
                     /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"
     regulatory      3584..3593
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4161..4282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4198..4217)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4252..4271
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4289..4744)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4426..4447)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4638..4657
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4771..4875
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4877..5234
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4877..5068
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4900..4920)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5085..5220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5269..5368,1..695)
                     /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             join(5269..5368,1..695)
                     /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(5323..5342)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
       61 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
      121 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
      181 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
      241 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
      301 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
      361 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
      421 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
      481 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
      541 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
      601 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
      661 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
      721 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
      781 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
      841 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
      901 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
      961 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     1021 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     1081 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     1141 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     1201 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     1261 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1321 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1381 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1441 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1501 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     1561 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     1621 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     1681 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     1741 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     1801 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     1861 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     1921 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     1981 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     2041 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     2101 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     2161 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     2221 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     2281 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     2341 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2401 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2461 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2521 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     2581 tgaaccgtca gatccgctag cgctaccgga ctcagatctc gagatgcagt gggccgtggg
     2641 ccggcggtgg gcgtgggccg cgctgctcct ggctgtcgca gcggtgctga cccaggtcgt
     2701 ctggctctgg ctgggtacgc agagcttcgt cttccagcgc gaagagatag cgcagttggc
     2761 gcggcagtac gctgggctgg accacgaact cgcatttagc agactgatcg tggagctgcg
     2821 gcggctgcac ccaggccacg tgctgcccga cgaggagctg cagtgggtgt tcgtcaacgc
     2881 tggtggatgg atgggcgcca tgtgccttct gcacgcctcg ctgtccgagt atgtgctgct
     2941 cttcggcacc gccttgggct cccgcggcca ctcggggcgc tactgggctg agatctcgga
     3001 taccatcatc tctggcacct tccaccagtg gagagagggc accaccaaaa gtgaggtctt
     3061 ctacccaggg gagacggtag tacacgggcc tggtgaggca acagctgtgg agtgggggcc
     3121 caatacttgg atggtcgagt acggccgggg cgtcatccca tccaccctgg ccttcgcgct
     3181 ggccgacact gtcttcagca cccaggacgc cctcaccgct gcatatactg ctcgctccgc
     3241 tgctcggggc gcacgcttgg aagctacgac agccctcttt ggccaggacc ctagggtacc
     3301 gcgggcccgg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagg ataacatggc
     3361 ctctctccca gcgacacatg agttacacat ctttggctcc atcaacggtg tggactttga
     3421 catggtgggt cagggcaccg gcaatccaaa tgatggttat gaggagttaa acctgaagtc
     3481 caccaagggt gacctccagt tctccccctg gattctggtc cctcatatcg ggtatggctt
     3541 ccatcagtac ctgccctacc ctgacgggat gtcgcctttc caggccgcca tggtagatgg
     3601 ctccggctac caagtccatc gcacaatgca gtttgaagat ggtgcctccc ttactgttaa
     3661 ctaccgctac acctacgagg gaagccacat caaaggagag gcccaggtga aggggactgg
     3721 tttccctgct gacggtcctg tgatgaccaa ctcgctgacc gctgcggact ggtgcaggtc
     3781 gaagaagact taccccaacg acaaaaccat catcagtacc tttaagtgga gttacaccac
     3841 tggaaatggc aagcgctacc ggagcactgc gcggaccacc tacacctttg ccaagccaat
     3901 ggcggctaac tatctgaaga accagccgat gtacgtgttc cgtaagacgg agctcaagca
     3961 ctccaagacc gagctcaact tcaaggagtg gcaaaaggcc tttaccgatg tgatgggcat
     4021 ggacgagctg tacaagtaag cggccgcgac tctagatcat aatcagccat accacatttg
     4081 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     4141 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     4201 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4261 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     4321 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4381 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4441 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4501 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4561 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4621 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     4681 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4741 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4801 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4861 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     4921 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4981 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5041 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5101 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5161 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5221 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5281 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5341 gcacaacaga caatcggctg ctctgatg
//
