LOCUS       sequence-451908-        5362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes siRNA resistant SigmaR1 protein labeled with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5362)
  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 5362)
  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-mCherry
FEATURES             Location/Qualifiers
     source          1..5362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(186..980)
                     /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(186..980)
                     /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(297..316)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     907..926
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1015..1372)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1029..1164
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1181..1372
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1329..1349
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(1374..1478)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1505..1960
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1592..1611)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1802..1823
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1967..2088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1978..1997)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2032..2051
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2210..2920)
                     /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(2488..2507)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2757..2775
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(3670..3873)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        3874..4177
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4352..4940)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4432..4451)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5269..5316
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5341..5360
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
ORIGIN
        1 tagcctcccc ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat
       61 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga
      121 atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt
      181 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
      241 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
      301 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
      361 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
      421 gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct
      481 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
      541 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
      601 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
      661 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
      721 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
      781 agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
      841 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
      901 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
      961 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca
     1021 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     1081 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     1141 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     1201 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     1261 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     1321 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc
     1381 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1441 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1501 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1561 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1621 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1681 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1741 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1801 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1861 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1921 tttaacgcga attttaacaa aatattaacg cttacaattt acgccttaag atacattgat
     1981 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2041 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2101 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     2161 aacctctaca aatgtggtat ggctgattat gatctagagt cgcggccgcc tacttgtaca
     2221 gctcgtccat gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg
     2281 tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt gacgttgtag gcgccgggca
     2341 gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt
     2401 ccttcagctt cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc
     2461 gctcggagga ggcctcccag cccatggtct tcttctgcat tacggggccg tcggagggga
     2521 agttggtgcc gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg gaggagtcct
     2581 gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac ttgaagccct
     2641 cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg taggccttgg
     2701 agccgtacat gaactgaggg gacaggatgt cccaggcgaa gggcaggggg ccacccttgg
     2761 tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc tcgccctcga
     2821 tctcgaactc gtggccgttc acggagccct ccatgtgcac cttgaagcgc atgaactcct
     2881 tgatgatggc catgttatcc tcctcgccct tgctcaccat gaccggtgga tcccgggccc
     2941 gcggtaccct agggtcctgg ccaaagaggt atgtcgtcaa ttccaagcgg aggccccgag
     3001 cataggagcg aagagtatag aagagggtga ggaagtcctg ggtgctgaag acagtgtcgg
     3061 ccagcgcgaa ggccagggtg gatgggatga cgccccggcc gtactcgacc atccaagtat
     3121 tgggccccca ctccacagct gttgcctcac caggcccgtg tactaccgtc tcccctgggt
     3181 agaagacctc acttttggtg gtgccctctc tccactggtg gaaggtgcca gagatgatgg
     3241 tatccgagat ctcagcccag tagcgccccg agtggccgcg ggagcccaag gcggtgccga
     3301 agagcagcac atactcggac agcgaggcgt gcagaaggca catggcgccc atccatccac
     3361 cagcgttgac gaacacccac tgcagctcct cgtcgggcag cacgtggcct gggtgcagcc
     3421 gccgcagctc cacgatcagt ctgctaaatg cgagttcgtg gtccagccca gcgtactgcc
     3481 gcgccaactg cgctatctct tcgcgctgga agacgaagct ctgcgtaccc agccagagcc
     3541 agacgacctg ggtcagcacc gctgcgacag ccaggagcag cgcggcccac gcccaccgcc
     3601 ggcccacggc ccactgcatc tcgagatctg agtccggtag cgctagcgga tctgacggtt
     3661 cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     3721 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     3781 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     3841 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     3901 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     3961 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     4021 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     4081 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     4141 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     4201 aactaatgac cccgtaattg attactatta ataactaatg catggcggta atacggttat
     4261 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4321 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4381 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4441 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4501 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4561 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4621 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4681 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4741 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4801 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4861 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4921 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4981 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5041 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg
     5101 aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac
     5161 ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg
     5221 gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc
     5281 caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta
     5341 ttgtctcctt ccgtgtttca gt
//
