LOCUS       sequence-263647-        6703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cry2PHR - mCherry - CofilinS3E protein fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6703)
  AUTHORS   Salem FB, Bunner WP, Prabhu VV, Kuyateh AB, O'Bryant CT, Murashov 
            AK, Szatmari EM, Hughes RM
  TITLE     CofActor: A light- and stress-gated optogenetic clustering tool to 
            study disease-associated cytoskeletal dynamics in living cells.
  JOURNAL   J Biol Chem. 2020 Aug 7;295(32):11231-11245. doi: 
            10.1074/jbc.RA119.012427. Epub 2020 May 18.
  PUBMED    32424038
REFERENCE   2  (bases 1 to 6703)
  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"; doi: 
            "10.1074/jbc.RA119.012427"; journalName: "J Biol Chem"; date: 
            "2020-08-7- 7"; volume: "295"; issue: "32"; pages: "11231-11245"
COMMENT     Sequence Label: Cry2PHR.mCh.CofS3E
FEATURES             Location/Qualifiers
     source          1..6703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        99..402
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        403..606
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     557..577
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             663..2156
                     /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
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     regulatory      2175..2184
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2181..2888
                     /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(2326..2344)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2594..2613
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3529..3650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3566..3585)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3620..3639
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3657..4112)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3794..3815)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4006..4025
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4139..4243
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4245..4602
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4245..4436
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4268..4288)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4453..4588
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4637..5431
                     /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             4637..5431
                     /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(4691..4710)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5301..5320
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5619..5638)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5663..5710
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6039..6627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6528..6547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
       61 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      121 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      181 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      241 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      301 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      361 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      421 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      481 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      541 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      601 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat ctcgaggcca
      661 ccatgaagat ggacaaaaag actatagttt ggtttagaag agacctaagg attgaggata
      721 atcctgcatt agcagcagct gctcacgaag gatctgtttt tcctgtcttc atttggtgtc
      781 ctgaagaaga aggacagttt tatcctggaa gagcttcaag atggtggatg aaacaatcac
      841 ttgctcactt atctcaatcc ttgaaggctc ttggatctga cctcacttta atcaaaaccc
      901 acaacacgat ttcagcgatc ttggattgta tccgcgttac cggtgctaca aaagtcgtct
      961 ttaaccacct ctatgatcct gtttcgttag ttcgggacca taccgtaaag gagaagctgg
     1021 tggaacgtgg gatctctgtg caaagctaca atggagatct attgtatgaa ccgtgggaga
     1081 tatactgcga aaagggcaaa ccttttacga gtttcaattc ttactggaag aaatgcttag
     1141 atatgtcgat tgaatccgtt atgcttcctc ctccttggcg gttgatgcca ataactgcag
     1201 cggctgaagc gatttgggcg tgttcgattg aagaactagg gctggagaat gaggccgaga
     1261 aaccgagcaa tgcgttgtta actagagctt ggtctccagg atggagcaat gctgataagt
     1321 tactaaatga gttcatcgag aagcagttga tagattatgc aaagaacagc aagaaagttg
     1381 ttgggaattc tacttcacta ctttctccgt atctccattt cggggaaata agcgtcagac
     1441 acgttttcca gtgtgcccgg atgaaacaaa ttatatgggc aagagataag aacagtgaag
     1501 gagaagaaag tgcagatctt tttcttaggg gaatcggttt aagagagtat tctcggtata
     1561 tatgtttcaa cttcccgttt actcacgagc aatcgttgtt gagtcatctt cggtttttcc
     1621 cttgggatgc tgatgttgat aagttcaagg cctggagaca aggcaggacc ggttatccgt
     1681 tggtggatgc cggaatgaga gagctttggg ctaccggatg gatgcataac agaataagag
     1741 tgattgtttc aagctttgct gtgaagtttc ttctccttcc atggaaatgg ggaatgaagt
     1801 atttctggga tacacttttg gatgctgatt tggaatgtga catccttggc tggcagtata
     1861 tctctgggag tatccccgat ggccacgagc ttgatcgctt ggacaatccc gcgttacaag
     1921 gcgccaaata tgacccagaa ggtgagtaca taaggcaatg gcttcccgag cttgcgagat
     1981 tgccaactga atggatccat catccatggg acgctccttt aaccgtactc aaagcttctg
     2041 gtgtggaact cggaacaaac tatgcgaaac ccattgtaga catcgacaca gctcgtgagc
     2101 tactagctaa agctatttca agaacccgtg aagcacagat catgatcgga gcagcagccc
     2161 gggatccacc ggtcgccacc atggtgagca agggcgagga ggataacatg gccatcatca
     2221 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
     2281 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     2341 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     2401 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     2461 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     2521 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     2581 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc
     2641 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     2701 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     2761 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     2821 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     2881 tgtacaaggg cggcagcggc ggcggcatgg ccgaaggtgt ggctgtctct gatggtgtca
     2941 tcaaggtgtt caatgacatg aaggttcgca agtcttcaac accagaagaa gtgaagaaac
     3001 gcaagaaggc ggtgctcttt tgcctgagtg aggacaagaa gaacatcatc ctggaggagg
     3061 gcaaggagat cctggtagga gatgtggggc agactgtgga cgacccctac accacttttg
     3121 tcaagatgct gccagacaag gactgccgct atgcactcta tgatgcaacc tatgagacca
     3181 aggagagcaa gaaggaggac ctggtgttca tcttctgggc ccccgagaat gcacccctca
     3241 agagcaaaat gatctatgcc agctccaagg atgccatcaa gaagaagctg acaggaatca
     3301 agcatgaatt acaagctaac tgctacgagg aggtcaagga ccgctgcacc ctggcagaga
     3361 aactaggtgg cagcgccgtc atttccctgg agggcaagcc tttgtgagcg gccgcgactc
     3421 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3481 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3541 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3601 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     3661 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3721 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3781 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3841 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3901 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3961 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4021 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4081 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     4141 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4201 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     4261 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4321 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4381 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4441 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4501 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4561 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     4621 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4681 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4741 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4801 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4861 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4921 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4981 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5041 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5101 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5161 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5221 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5281 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5341 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5401 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5461 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5521 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5581 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     5641 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     5701 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5761 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5821 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5881 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5941 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6001 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6061 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6121 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6181 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6241 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6301 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6361 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6421 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6481 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6541 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6601 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6661 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tat
//
