LOCUS       sequence-442209-        6406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CRY2olig-mCherry fused to a codon-optimised poly-GR 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6406)
  AUTHORS   Hodgson RE, Rayment JA, Huang WP, Sanchez Avila A, Ellis BCS, Lin 
            YH, Soni N, Hautbergue GM, Shelkovnikova TA
  TITLE     C9orf72 poly-PR forms anisotropic condensates causative of nuclear 
            TDP-43 pathology.
  JOURNAL   iScience. 2024 Sep 14;27(10):110937. doi: 
            10.1016/j.isci.2024.110937. eCollection 2024 Oct 18.
  PUBMED    39391721
REFERENCE   2  (bases 1 to 6406)
  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: "iScience.";
            date: "2024-09-14"; volume: "27(10):110937. doi"; pages: " 
            10.1016/j.isci.2024.110937. eCollection 2024 Oct 18"
COMMENT     Sequence Label: CRY2olig-GR
FEATURES             Location/Qualifiers
     source          1..6406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        87..444
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        87..278
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(110..130)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      295..430
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             479..1273
                     /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             479..1273
                     /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(533..552)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1143..1162
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1461..1480)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1505..1552
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1881..2469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2370..2389
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2644..2947
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2948..3151
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3102..3122
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3208..4701
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTRGAQIMIGAA"
     regulatory      4720..4729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4726..5433
                     /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(4871..4889)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5139..5158
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5777..5898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5814..5833)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5868..5887
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5905..6360)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6042..6063)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6254..6273
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6387..6406,1..85)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
       61 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
      121 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
      181 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      241 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
      301 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
      361 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
      421 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
      481 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
      541 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
      601 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
      661 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
      721 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
      781 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
      841 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
      901 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
      961 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     1021 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     1081 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     1141 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     1201 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     1261 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     1321 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     1381 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     1441 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     1501 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     1561 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     1621 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     1681 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     1741 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1801 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1861 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1921 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1981 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2041 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2101 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2161 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2221 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2281 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2341 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2401 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2461 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2521 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     2581 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     2641 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     2701 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     2761 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     2821 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     2881 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     2941 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     3001 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     3061 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     3121 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
     3181 ctaccggact cagatctcga ggccaccatg aagatggaca aaaagactat agtttggttt
     3241 agaagagacc taaggattga ggataatcct gcattagcag cagctgctca cgaaggatct
     3301 gtttttcctg tcttcatttg gtgtcctgaa gaagaaggac agttttatcc tggaagagct
     3361 tcaagatggt ggatgaaaca atcacttgct cacttatctc aatccttgaa ggctcttgga
     3421 tctgacctca ctttaatcaa aacccacaac acgatttcag cgatcttgga ttgtatccgc
     3481 gttaccggtg ctacaaaagt cgtctttaac cacctctatg atcctgtttc gttagttcgg
     3541 gaccataccg taaaggagaa gctggtggaa cgtgggatct ctgtgcaaag ctacaatgga
     3601 gatctattgt atgaaccgtg ggagatatac tgcgaaaagg gcaaaccttt tacgagtttc
     3661 aattcttact ggaagaaatg cttagatatg tcgattgaat ccgttatgct tcctcctcct
     3721 tggcggttga tgccaataac tgcagcggct gaagcgattt gggcgtgttc gattgaagaa
     3781 ctagggctgg agaatgaggc cgagaaaccg agcaatgcgt tgttaactag agcttggtct
     3841 ccaggatgga gcaatgctga taagttacta aatgagttca tcgagaagca gttgatagat
     3901 tatgcaaaga acagcaagaa agttgttggg aattctactt cactactttc tccgtatctc
     3961 catttcgggg aaataagcgt cagacacgtt ttccagtgtg cccggatgaa acaaattata
     4021 tgggcaagag ataagaacag tgaaggagaa gaaagtgcag atctttttct taggggaatc
     4081 ggtttaagag agtattctcg gtatatatgt ttcaacttcc cgtttactca cgagcaatcg
     4141 ttgttgagtc atcttcggtt tttcccttgg gatgctgatg ttgataagtt caaggcctgg
     4201 agacaaggca ggaccggtta tccgttggtg gatgccggaa tgagagagct ttgggctacc
     4261 ggatggatgc ataacagaat aagagtgatt gtttcaagct ttgctgtgaa gtttcttctc
     4321 cttccatgga aatggggaat gaagtatttc tgggatacac ttttggatgc tgatttggaa
     4381 tgtgacatcc ttggctggca gtatatctct gggagtatcc ccgatggcca cgagcttgat
     4441 cgcttggaca atcccgcgtt acaaggcgcc aaatatgacc cagaaggtga gtacataagg
     4501 caatggcttc ccgagcttgc gagattgcca actgaatgga tccatcatcc atgggacgct
     4561 cctttaaccg tactcaaagc ttctggtgtg gaactcggaa caaactatgc gaaacccatt
     4621 gtagacatcg acacagctcg tgagctacta gctaaagcta tttcaagaac ccgtggagca
     4681 cagatcatga tcggagcagc agcccgggat ccaccggtcg ccaccatggt gagcaagggc
     4741 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     4801 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     4861 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     4921 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     4981 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     5041 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     5101 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     5161 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     5221 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     5281 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     5341 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     5401 cactccaccg gcggcatgga cgagctgtac aagatgggca gaggccgggg aagaggcaga
     5461 ggacgtggtc gaggacgtgg caggggcaga ggtagaggaa ggggacgagg acgcggaaga
     5521 ggaagagggc gaggccgagg caggggacgc ggccgagggc gaggaagagg acggggtaga
     5581 gggcggggac gtggaagggg aaggggacgg ggcaggggac gaggcagagg ccgcggacgt
     5641 ggtagaggca gataagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     5701 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5761 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5821 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5881 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     5941 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     6001 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6061 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6121 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     6181 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6241 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6301 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6361 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgttt
//
