LOCUS       sequence-451919-        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 a4mut (I178...
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    291..412
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(328..347)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     382..401
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(419..874)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(556..577)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     768..787
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        901..1005
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1007..1364
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1007..1198
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1030..1050)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1215..1350
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1399..2193
                     /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             1399..2193
                     /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     2063..2082
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2381..2400)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2425..2472
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2801..3389
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3290..3309
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3564..3867
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3868..4071
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      4821..4830
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4827..5368,1..169)
                     /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      5082..5091
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gcggaccacc tacacctttg ccaagccaat ggcggctaac tatctgaaga accagccgat
       61 gtacgtgttc cgtaagacgg agctcaagca ctccaagacc gagctcaact tcaaggagtg
      121 gcaaaaggcc tttaccgatg tgatgggcat ggacgagctg tacaagtaag cggccgcgac
      181 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
      241 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
      301 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
      361 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
      421 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
      481 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
      541 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
      601 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
      661 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
      721 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
      781 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
      841 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
      901 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      961 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     1021 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     1081 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     1141 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     1201 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     1261 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     1321 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     1381 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     1441 tgggtggaga ggctattcgg mtatgactgg gcacaacaga caatcggctg ctctgatgcc
     1501 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     1561 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     1621 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     1681 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     1741 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     1801 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     1861 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     1921 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     1981 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     2041 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     2101 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     2161 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     2221 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     2281 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     2341 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     2401 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     2461 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     2521 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     2581 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     2641 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2701 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2761 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2821 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2881 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2941 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3001 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3061 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3121 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3181 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3241 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3301 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3361 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3421 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3481 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     3541 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     3601 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     3661 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     3721 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     3781 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     3841 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     3901 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     3961 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     4021 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     4081 aaccgtcaga tccgctagcg ctaccggact cagatctcga gatgcagtgg gccgtgggcc
     4141 ggcggtgggc gtgggccgcg ctgctcctgg ctgtcgcagc ggtgctgacc caggtcgtct
     4201 ggctctggct gggtacgcag agcttcgtct tccagcgcga agagatagcg cagttggcgc
     4261 ggcagtacgc tgggctggac cacgaactcg catttagcag actgatcgtg gagctgcggc
     4321 ggctgcaccc aggccacgtg ctgcccgacg aggagctgca gtgggtgttc gtcaacgctg
     4381 gtggatggat gggcgccatg tgccttctgc acgcctcgct gtccgagtat gtgctgctct
     4441 tcggcaccgc cttgggctcc cgcggccact cggggcgcta ctgggctgag atctcggata
     4501 ccatcatctc tggcaccttc caccagtgga gagagggcac caccaaaagt gaggtcttct
     4561 acccagggga gacggtagta cacgggcctg gtgaggcaac agctgtggag tgggggccca
     4621 atacttggat ggtcgagtac ggccggggcg tcgcgccatc caccgcagcc ttcgcggctg
     4681 ccgacactgc cttcagcacc caggacttcc tcaccctctt ctatactctt cgctcctatg
     4741 ctcggggcct ccgcttggaa ttgacgacat acctctttgg ccaggaccct agggtaccgc
     4801 gggcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggaggat aacatggcct
     4861 ctctcccagc gacacatgag ttacacatct ttggctccat caacggtgtg gactttgaca
     4921 tggtgggtca gggcaccggc aatccaaatg atggttatga ggagttaaac ctgaagtcca
     4981 ccaagggtga cctccagttc tccccctgga ttctggtccc tcatatcggg tatggcttcc
     5041 atcagtacct gccctaccct gacgggatgt cgcctttcca ggccgccatg gtagatggct
     5101 ccggctacca agtccatcgc acaatgcagt ttgaagatgg tgcctccctt actgttaact
     5161 accgctacac ctacgaggga agccacatca aaggagaggc ccaggtgaag gggactggtt
     5221 tccctgctga cggtcctgtg atgaccaact cgctgaccgc tgcggactgg tgcaggtcga
     5281 agaagactta ccccaacgac aaaaccatca tcagtacctt taagtggagt tacaccactg
     5341 gaaatggcaa gcgctaccgg agcactgc
//
