LOCUS       sequence-442218-        7765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CRY2olig-mCherry fused to FUS .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7765)
  TITLE     Expression of CRY2olig-mCherry fusions
REFERENCE   2  (bases 1 to 7765)
  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"
COMMENT     Sequence Label: CRY2olig-FUS
FEATURES             Location/Qualifiers
     source          1..7765
                     /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..5430
                     /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
                     EGRHSTGGMDELY"
     primer_bind     complement(4871..4889)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5139..5158
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5814..5825
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    7136..7257
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7173..7192)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7227..7246
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(7264..7719)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7401..7422)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7613..7632
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(7746..7765,1..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 gagatggcct caaacgatta tacccaacaa
     5461 gcaacccaaa gctatggggc ctaccccacc cagcccgggc agggctattc ccagcagagc
     5521 agtcagccct acggacagca gagttacagt ggttatagcc agtccacgga cacttcaggc
     5581 tatggccaga gcagctattc ttcttatggc cagagccaga acacaggcta tggaactcag
     5641 tcaactcccc agggatatgg ctcgactggc ggctatggca gtagccagag ctcccaatcg
     5701 tcttacgggc agcagtcctc ctatcctggc tatggccagc agccagctcc cagcagcacc
     5761 tcgggaagtt acggtagcag ttctcagagc agcagctatg ggcagcccca gagtgggagc
     5821 tacagccagc agcctagcta tggtggacag cagcaaagct atggacagca gcaaagctat
     5881 aatccccctc agggctatgg acagcagaac cagtacaaca gcagcagtgg tggtggaggt
     5941 ggaggtggag gtggaggtaa ctatggccaa gatcaatcct ccatgagtag tggtggtggc
     6001 agtggtggcg gttatggcaa tcaagaccag agtggtggag gtggcagcgg tggctatgga
     6061 cagcaggacc gtggaggccg cggcaggggt ggcagtggtg gcggcggcgg cggcggcggt
     6121 ggtggttaca accgcagcag tggtggctat gaacccagag gtcgtggagg tggccgtgga
     6181 ggcagaggtg gcatgggcgg aagtgaccgt ggtggcttca ataaatttgg tggccctcgg
     6241 gaccaaggat cacgtcatga ctccgaacag gataattcag acaacaacac catctttgtg
     6301 caaggcctgg gtgagaatgt tacaattgag tctgtggctg attacttcaa gcagattggt
     6361 attattaaga caaacaagaa aacgggacag cccatgatta atttgtacac agacagggaa
     6421 actggcaagc tgaagggaga ggcaacggtc tcttttgatg acccaccttc agctaaagca
     6481 gctattgact ggtttgatgg taaagaattc tccggaaatc ctatcaaggt ctcatttgct
     6541 actcgccggg cagactttaa tcggggtggt ggcaatggtc gtggaggccg agggcgagga
     6601 ggacccatgg gccgtggagg ctatggaggt ggtggcagtg gtggtggtgg ccgaggagga
     6661 tttcccagtg gaggtggtgg cggtggagga cagcagcgag ctggtgactg gaagtgtcct
     6721 aatcccacct gtgagaatat gaacttctct tggaggaatg aatgcaacca gtgtaaggcc
     6781 cctaaaccag atggcccagg agggggacca ggtggctctc acatgggggg taactacggg
     6841 gatgatcgtc gtggtggcag aggaggctat gatcgaggcg gctaccgggg ccgcggcggg
     6901 gaccgtggag gcttccgagg gggccggggt ggtggggaca gaggtggctt tggccctggc
     6961 aagatggatt ccaggggtga gcacagacag gatcgcaggg agaggccgta ttaagcggcc
     7021 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     7081 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     7141 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     7201 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     7261 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     7321 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     7381 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     7441 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     7501 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     7561 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     7621 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     7681 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     7741 atgtgcgcgg aacccctatt tgttt
//
