LOCUS       sequence-260103-        6327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stress granule marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6327)
  AUTHORS   Yaginuma H,  Okada Y
  TITLE     Live cell imaging of metabolic heterogeneity by quantitative 
            fluorescent ATP indicator protein, QUEEN-37C
  JOURNAL   bioRxiv 2021.10.08.463131
REFERENCE   2  (bases 1 to 6327)
  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: "bioRxiv 
            2021.10.08.463131"
COMMENT     Sequence Label: pN1/G3BP1-iRFP
FEATURES             Location/Qualifiers
     source          1..6327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    3055..3176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3092..3111)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3146..3165
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3183..3638)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3320..3341)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3532..3551
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3665..3769
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3771..4128
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3771..3962
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3794..3814)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3979..4114
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4163..4957
                     /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             4163..4957
                     /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(4217..4236)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4827..4846
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5145..5164)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5189..5236
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5565..6153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6054..6073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcca ccatggtgat ggagaagcct agtcccctgc
      601 tggtcgggcg ggaatttgtg agacagtatt acacactgct gaaccaggcc ccagacatgc
      661 tgcatagatt ttatggaaag aactcttctt atgtccatgg gggattggat tcaaatggaa
      721 agccagcaga tgcagtctac ggacagaaag aaatccacag gaaagtgatg tcacaaaact
      781 tcaccaactg ccacaccaag attcgccatg ttgatgctca tgccacgcta aatgatggtg
      841 tggtagtcca ggtgatgggg cttctctcta acaacaacca ggctttgagg agattcatgc
      901 aaacgtttgt ccttgctcct gaggggtctg ttgcaaataa attctatgtt cacaatgata
      961 tcttcagata ccaagatgag gtctttggtg ggtttgtcac tgagcctcag gaggagtctg
     1021 aagaagaagt agaggaacct gaagaaagac agcaaacacc tgaggtggta cctgatgatt
     1081 ctggaacttt ctatgatcag gcagttgtca gtaatgacat ggaagaacat ttagaggagc
     1141 ctgttgctga accagagcct gatcctgaac cagaaccaga acaagaacct gtatctgaaa
     1201 tccaagagga aaagcctgag ccagtattag aagaaactgc ccctgaggat gctcagaaga
     1261 gttcttctcc agcacctgca gacatagctc agacagtaca ggaagacttg aggacatttt
     1321 cttgggcatc tgtgaccagt aagaatcttc cacccagtgg agctgttcca gttactggga
     1381 taccacctca tgttgttaaa gtaccagctt cacagccccg tccagagtct aagcctgaat
     1441 ctcagattcc accacaaaga cctcagcggg atcaaagagt gcgagaacaa cgaataaata
     1501 ttcctcccca aaggggaccc agaccaatcc gtgaggctgg tgagcaaggt gacattgaac
     1561 cccgaagaat ggtgagacac cctgacagtc accaactctt cattggcaac ctgcctcatg
     1621 aagtggacaa atcagagctt aaagatttct ttcaaagtta tggaaacgtg gtggagttgc
     1681 gcattaacag tggtgggaaa ttacccaatt ttggttttgt tgtgtttgat gattctgagc
     1741 ctgttcagaa agtccttagc aacaggccca tcatgttcag aggtgaggtc cgtctgaatg
     1801 tcgaagagaa gaagactcga gctgccaggg aaggcgaccg acgagataat cgccttcggg
     1861 gacctggagg ccctcgaggt gggctgggtg gtggaatgag aggccctccc cgtggaggca
     1921 tggtgcagaa accaggattt ggagtgggaa gggggcttgc gccacggcag cgggatccac
     1981 cggtcgccga agggagcgtg gcaagacagc ccgatctgct gacatgcgac gacgaaccaa
     2041 tccacattcc aggagctatc cagcctcatg gactgctgct ggctctggcc gctgacatga
     2101 ctatcgtggc aggctctgat aacctgccag agctgaccgg gctggctatc ggagcactga
     2161 ttggccggtc cgcagccgac gtgttcgatt ctgagactca caatagactg accatcgccc
     2221 tggctgaacc tggggctgca gtgggagccc caattaccgt cggcttcaca atgcgaaagg
     2281 acgctggctt tatcgggtcc tggcaccgcc atgatcagct gatttttctg gagctggaac
     2341 cacctcagag agacgtggca gagccacagg ctttctttcg gagaacaaac agcgccatca
     2401 ggcgcctgca ggccgctgag actctggaat ccgcttgcgc agccgctgca caggaagtga
     2461 ggaagatcac cgggttcgat cgcgtcatga tctacagatt cgcctcagac tttagcggag
     2521 aagtgatcgc agaagatcgc tgtgccgagg tcgaaagtaa actgggcctg cattacccag
     2581 cctcaaccgt gcctgcacag gcacgacggc tgtatacaat caaccccgtg cggatcattc
     2641 ccgacattaa ttacagacca gtgcccgtca cacctgacct gaacccagtg actgggaggc
     2701 ccatcgatct gagcttcgcc attctgcgca gcgtgtcccc tgtccacctg gagtttatga
     2761 ggaatatcgg aatgcatggc accatgtcta tcagtattct gagaggcgaa aggctgtggg
     2821 ggctgatcgt gtgccaccat cggacacctt actatgtgga cctggacggc aggcaggctt
     2881 gtgaactggt ggctcaggtg ctggcttggc agattggcgt gatggaagaa tgagcggccg
     2941 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3001 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3061 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3121 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     3181 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3241 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3301 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3361 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3421 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3481 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3541 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3601 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3661 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3721 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3781 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3841 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3901 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3961 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4021 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4081 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4141 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4201 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4261 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4321 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4381 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4441 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4501 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4561 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4621 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4681 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4741 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4801 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4861 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4921 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4981 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5041 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5101 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5161 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5221 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5281 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5341 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5401 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5461 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5521 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5581 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5641 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5701 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5761 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5821 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5881 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5941 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6001 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6061 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6121 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6181 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6241 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     6301 agttcatagc ccatatatgg agttccg
//
