LOCUS       sequence-451901-        5281 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes mutant SigmaR1 protein (transmembrane region deletion) 
            labeled with mNeonGreen .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5281)
  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 5281)
  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: ss-SigmaR1DTM-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..5281
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      282..291
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      543..552
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1215..1418)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1244..1264)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1419..1722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1897..2485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1977..1996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2814..2861
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2886..2905
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3093..3887)
                     /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(3093..3887)
                     /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(3204..3223)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3814..3833
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3922..4279)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3936..4071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4088..4279
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4236..4256
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4281..4385)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4412..4867
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4499..4518)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4709..4730
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4874..4995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4885..4904)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4939..4958
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5117..5281,1..546))
                     /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 ccgcgcagtg ctccggtagc gcttgccatt tccagtggtg taactccact taaaggtact
       61 gatgatggtt ttgtcgttgg ggtaagtctt cttcgacctg caccagtccg cagcggtcag
      121 cgagttggtc atcacaggac cgtcagcagg gaaaccagtc cccttcacct gggcctctcc
      181 tttgatgtgg cttccctcgt aggtgtagcg gtagttaaca gtaagggagg caccatcttc
      241 aaactgcatt gtgcgatgga cttggtagcc ggagccatct accatggcgg cctggaaagg
      301 cgacatcccg tcagggtagg gcaggtactg atggaagcca tacccgatat gagggaccag
      361 aatccagggg gagaactgga ggtcaccctt ggtggacttc aggtttaact cctcataacc
      421 atcatttgga ttgccggtgc cctgacccac catgtcaaag tccacaccgt tgatggagcc
      481 aaagatgtgt aactcatgtg tcgctgggag agaggccatg ttatcctcct cgcccttgct
      541 caccatggtg gcgaccggtg gatcccgggc ccgcggtacc ctagggtcct ggccaaagag
      601 gtaggtggtg agctcaagcc ggaggccccg agcataggag cgaagagtat agaagagggt
      661 gaggaagtcc tgggtgctga agacagtgtc ggccagcgcg aaggccaggg tggatgggat
      721 gacgccccgg ccgtactcca ccatccatgt gtttggcccc cactccacag ctgttgcctc
      781 accaggcccg tgtactaccg tctcccctgg gtagaagacc tcacttttgg tggtgccctc
      841 tctccactgg tggaaggtgc cagagatgat ggtatccgag atctcagccc agtagcgccc
      901 cgagtggccg cgggagccca aggcggtgcc gaagagcagc acatactcgg acagcgaggc
      961 gtgcagaagg cacatggcgc ccatccagcc acccgcattc acgaacaccc actgcagctc
     1021 ctcgtcgggc agcacgtggc ctgggtgcag ccgccgcagc tccacgatca gacgagagaa
     1081 ggccagctcg tggtccagcc cagcgtactg ccgcgccaac tgcgctatct cttcgcgctg
     1141 gaagacgaag ctctgcgtac ccatctcgag atctgagtcc ggtagcgcta gcggatctga
     1201 cggttcacta aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt
     1261 tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa
     1321 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
     1381 tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
     1441 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     1501 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     1561 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     1621 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     1681 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     1741 ctatgaacta atgaccccgt aattgattac tattaataac taatgcatgg cggtaatacg
     1801 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1861 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1921 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1981 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2041 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2101 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2161 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2221 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2281 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2341 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2401 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2461 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2521 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2581 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2641 acctgaggct atggcagggc ctgccgcccc gacgttggct gcgagccctg ggccttcacc
     2701 cgaacttggg gggtggggtg gggaaaagga agaaacgcgg gcgtattggc cccaatgggg
     2761 tctcggtggg gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa
     2821 cgacccaaca ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc
     2881 cggtattgtc tccttccgtg tttcagttag cctcccccta gggtgggcga agaactccag
     2941 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
     3001 caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc
     3061 ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     3121 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     3181 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     3241 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     3301 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc
     3361 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     3421 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     3481 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     3541 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     3601 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     3661 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg
     3721 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     3781 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     3841 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     3901 catcctgtct cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     3961 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     4021 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     4081 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     4141 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     4201 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     4261 ctggttcttt ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt
     4321 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4381 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     4441 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4501 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4561 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4621 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4681 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4741 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4801 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     4861 acaatttacg ccttaagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     4921 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4981 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     5041 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     5101 ctagagtcgc ggccgcttac ttgtacagct cgtccatgcc catcacatcg gtaaaggcct
     5161 tttgccactc cttgaagttg agctcggtct tggagtgctt gagctccgtc ttacggaaca
     5221 cgtacatcgg ctggttcttc agatagttag ccgccattgg cttggcaaag gtgtaggtgg
     5281 t
//
