LOCUS       sequence-197490-        8851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fusion of CRY2low mutant (CRY2 aa1-491 R489E A491D) with 
            tdTomato and Raf1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8851)
  AUTHORS   Duan L, Hope J, Ong Q, Lou HY, Kim N, McCarthy C, Acero V, Lin MZ, 
            Cui B
  TITLE     Understanding CRY2 interactions for optical control of intracellular
            signaling.
  JOURNAL   Nat Commun. 2017 Sep 15;8(1):547. doi: 10.1038/s41467-017-00648-8.
  PUBMED    28916751
REFERENCE   2  (bases 1 to 8851)
  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: "Nat 
            Commun."; date: "2017-09-15"; volume: "8(1):547. doi"; pages: " 
            10.1038/s41467-017-00648-8"
COMMENT     Sequence Label: CRY2low-tdTomato-Raf1
FEATURES             Location/Qualifiers
     source          1..8851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2116..2125
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2122..3549
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     polyA_signal    5639..5760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5676..5695)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5730..5749
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5767..6222)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5904..5925)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6116..6135
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6249..6353
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6355..6712
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6355..6546
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6378..6398)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6563..6698
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6747..7541
                     /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             6747..7541
                     /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(6801..6820)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7411..7430
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7729..7748)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7773..7820
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8149..8737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8638..8657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc caccatgaag atggacaaaa agactatagt ttggtttaga
      661 agagacctaa ggattgagga taatcctgca ttagcagcag ctgctcacga aggatctgtt
      721 tttcctgtct tcatttggtg tcctgaagaa gaaggacagt tttatcctgg aagagcttca
      781 agatggtgga tgaaacaatc acttgctcac ttatctcaat ccttgaaggc tcttggatct
      841 gacctcactt taatcaaaac ccacaacacg atttcagcga tcttggattg tatccgcgtt
      901 accggtgcta caaaagtcgt ctttaaccac ctctatgatc ctgtttcgtt agttcgggac
      961 cataccgtaa aggagaagct ggtggaacgt gggatctctg tgcaaagcta caatggagat
     1021 ctattgtatg aaccgtggga gatatactgc gaaaagggca aaccttttac gagtttcaat
     1081 tcttactgga agaaatgctt agatatgtcg attgaatccg ttatgcttcc tcctccttgg
     1141 cggttgatgc caataactgc agcggctgaa gcgatttggg cgtgttcgat tgaagaacta
     1201 gggctggaga atgaggccga gaaaccgagc aatgcgttgt taactagagc ttggtctcca
     1261 ggatggagca atgctgataa gttactaaat gagttcatcg agaagcagtt gatagattat
     1321 gcaaagaaca gcaagaaagt tgttgggaat tctacttcac tactttctcc gtatctccat
     1381 ttcggggaaa taagcgtcag acacgttttc cagtgtgccc ggatgaaaca aattatatgg
     1441 gcaagagata agaacagtga aggagaagaa agtgcagatc tttttcttag gggaatcggt
     1501 ttaagagagt attctcggta tatatgtttc aacttcccgt ttactcacga gcaatcgttg
     1561 ttgagtcatc ttcggttttt cccttgggat gctgatgttg ataagttcaa ggcctggaga
     1621 caaggcagga ccggttatcc gttggtggat gccggaatga gagagctttg ggctaccgga
     1681 tggatgcata acagaataag agtgattgtt tcaagctttg ctgtgaagtt tcttctcctt
     1741 ccatggaaat ggggaatgaa gtatttctgg gatacacttt tggatgctga tttggaatgt
     1801 gacatccttg gctggcagta tatctctggg agtatccccg atggccacga gcttgatcgc
     1861 ttggacaatc ccgcgttaca aggcgccaaa tatgacccag aaggtgagta cataaggcaa
     1921 tggcttcccg agcttgcgag attgccaact gaatggatcc atcatccatg ggacgctcct
     1981 ttaaccgtac tcaaagcttc tggtgtggaa ctcggaacaa actatgcgaa acccattgta
     2041 gacatcgaca cagctcgtga gctactagct aaagctattt caagaaccga ggaagacgcc
     2101 cgggataagg atcccgccac catggtgagc aagggcgagg aggtcatcaa agagttcatg
     2161 cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag
     2221 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc
     2281 cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac
     2341 gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag
     2401 tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc
     2461 ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac
     2521 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc
     2581 cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac
     2641 tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac
     2701 tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
     2761 cagtacgagc gctccgaggg ccgccaccac ctgttcctgg ggcatggcac cggcagcacc
     2821 ggcagcggca gctccggcac cgcctcctcc gaggacaaca acatggccgt catcaaagag
     2881 ttcatgcgct tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag
     2941 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     3001 ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag
     3061 gcgtacgtga agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc
     3121 ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac
     3181 tcctccctgc aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc
     3241 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg
     3301 tacccccgcg acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc
     3361 ggccactacc tggtggagtt caagaccatc tacatggcca agaagcccgt gcaactgccc
     3421 ggctactact acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc
     3481 gtggaacagt acgagcgctc cgagggccgc caccacctgt tcctgtacgg catggacgag
     3541 ctgtacaagt ccggactcag atctcgagtg atggagcaca tacagggagc ttggaagacg
     3601 atcagcaatg gttttggatt caaagatgcc gtgtttgatg gctccagctg catctctcct
     3661 acaatagttc agcagtttgg ctatcagcgc cgggcatcag atgatggcaa actcacagat
     3721 ccttctaaga caagcaacac tatccgtgtt ttcttgccga acaagcaaag aacagtggtc
     3781 aatgtgcgaa atggaatgag cttgcatgac tgccttatga aagcactcaa ggtgaggggc
     3841 ctgcaaccag agtgctgtgc agtgttcaga cttctccacg aacacaaagg taaaaaagca
     3901 cgcttagatt ggaatactga tgctgcgtct ttgattggag aagaacttca agtagatttc
     3961 ctggatcatg ttcccctcac aacacacaac tttgctcgga agacgttcct gaagcttgcc
     4021 ttctgtgaca tctgtcagaa attcctgctc aatggatttc gatgtcagac ttgtggctac
     4081 aaatttcatg agcactgtag caccaaagta cctactatgt gtgtggactg gagtaacatc
     4141 agacaactct tattgtttcc aaattccact attggtgata gtggagtccc agcactacct
     4201 tctttgacta tgcgtcgtat gcgagagtct gtttccagga tgcctgttag ttctcagcac
     4261 agatattcta cacctcacgc cttcaccttt aacacctcca gtccctcatc tgaaggttcc
     4321 ctctcccaga ggcagaggtc gacatccaca cctaatgtcc acatggtcag caccaccctg
     4381 cctgtggaca gcaggatgat tgaggatgca attcgaagtc acagcgaatc agcctcacct
     4441 tcagccctgt ccagtagccc caacaatctg agcccaacag gctggtcaca gccgaaaacc
     4501 cccgtgccag cacaaagaga gcgggcacca gtatctggga cccaggagaa aaacaaaatt
     4561 aggcctcgtg gacagagaga ttcaagctat tattgggaaa tagaagccag tgaagtgatg
     4621 ctgtccactc ggattgggtc aggctctttt ggaactgttt ataagggtaa atggcacgga
     4681 gatgttgcag taaagatcct aaaggttgtc gacccaaccc cagagcaatt ccaggccttc
     4741 aggaatgagg tggctgttct gcgcaaaaca cggcatgtga acattctgct tttcatgggg
     4801 tacatgacaa aggacaacct ggcaattgtg acccagtggt gcgagggcag cagcctctac
     4861 aaacacctgc atgtccagga gaccaagttt cagatgttcc agctaattga cattgcccgg
     4921 cagacggctc agggaatgga ctatttgcat gcaaagaaca tcatccatag agacatgaaa
     4981 tccaacaata tatttctcca tgaaggctta acagtgaaaa ttggagattt tggtttggca
     5041 acagtaaagt cacgctggag tggttctcag caggttgaac aacctactgg ctctgtcctc
     5101 tggatggccc cagaggtgat ccgaatgcag gataacaacc cattcagttt ccagtcggat
     5161 gtctactcct atggcatcgt attgtatgaa ctgatgacgg gggagcttcc ttattctcac
     5221 atcaacaacc gagatcagat catcttcatg gtgggccgag gatatgcctc cccagatctt
     5281 agtaagctat ataagaactg ccccaaagca atgaagaggc tggtagctga ctgtgtgaag
     5341 aaagtaaagg aagagaggcc tctttttccc cagatcctgt cttccattga gctgctccaa
     5401 cactctctac cgaagatcaa ccggagcgct tccgagccat ccttgcatcg ggcagcccac
     5461 actgaggata tcaatgcttg cacgctgacc acgtccccga ggctgcctgt cttctaagcg
     5521 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     5581 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     5641 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5701 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5761 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     5821 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     5881 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     5941 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     6001 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     6061 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     6121 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     6181 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     6241 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6301 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     6361 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     6421 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     6481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     6541 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     6601 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     6661 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     6721 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     6781 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     6841 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     6901 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     6961 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     7021 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     7081 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     7141 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     7201 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     7261 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     7321 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     7381 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     7441 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     7501 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     7561 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     7621 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     7681 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     7741 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     7801 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     7861 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     7921 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     7981 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     8041 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     8101 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     8161 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     8221 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     8281 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     8341 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     8401 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     8461 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     8521 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     8581 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     8641 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     8701 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8761 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     8821 ttctgtggat aaccgtatta ccgccatgca t
//
