LOCUS       sequence-415636-        8109 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  A red fluorescent protein with improved monomericity enables 
            ratiometric voltage imaging with ASAP3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8109)
  AUTHORS   Kim BB, Wu H, Hao YA, Pan M, Chavarha M, Zhao Y, Westberg M, 
            St-Pierre F, Wu JC, Lin MZ
  TITLE     A red fluorescent protein with improved monomericity enables 
            ratiometric voltage imaging with ASAP3.
  JOURNAL   Sci Rep. 2022 Mar 7;12(1):3678. doi: 10.1038/s41598-022-07313-1.
  PUBMED    35256624
REFERENCE   2  (bases 1 to 8109)
  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: "Sci Rep."; 
            date: "2022-03-7"; volume: "12(1):3678. doi"; pages: " 
            10.1038/s41598-022-07313-1"
COMMENT     Sequence Label: pc3.1-CAGGS-ASAP3-mCyRFP3
FEATURES             Location/Qualifiers
     source          1..8109
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          105..1121
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1045..1067
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1129..1148
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1287..1296
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2598..3299
                     /codon_start=1
                     /product="monomeric cyan-excitable red fluorescent protein 
                     derived from mCyRFP1 (Kim et al., 2022)"
                     /label=mCyRFP3
                     /translation="MVSKGEELIKENMRSKLYLEGSVNGHQFKCTHEGEGKPYEGKQTA
                     RIKVVEGGPLPFAFDILATMFMYGSKVFIKYPADLPDYFKQSFPEGFTWERVMVFEDGG
                     VLTATQDTSLQDGELIYNVKLRGVNFPANGPVMQKKTLGWEPSTETMTPADGGLEGRCD
                     KVLKLVGGGHLHVNFKTTYKSKKPVKMPGVHYVDRRLERIKEADNETYVEQYEHAVARY
                     SNLGGGMDELYK"
     primer_bind     complement(3416..3433)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3422..3646
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3692..4120
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3779..3798)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3989..4010
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4129..4149)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4134..4463
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4314..4449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4376..4395
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4474..5073
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4474..4493)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4970..4990
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5233..5354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5270..5289)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5324..5343
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5403..5419)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5403..5419)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5416..5438)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5427..5443
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5451..5457,5458..5475,5476..5481))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5496..5517
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5634..5651)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5805..6393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5885..5904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6564..7355,7356..7424))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7187..7206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7425..7529)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7561..7940
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 grggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
       61 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
      121 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
      181 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
      241 gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc
      301 gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
      361 ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg
      421 ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
      481 gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag
      541 cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg
      601 ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg
      661 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cckcgggccg
      721 gggrgggctc gggggrgggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg
      781 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
      841 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
      901 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
      961 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc
     1021 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     1081 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     1141 ttattgtgct gtctcatcat tttggcaaag aattcctcga ggaattcact cctcaggtgc
     1201 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     1261 gagatctgca tgagctagct gtacacgcca ccatggagac gactgtgagg tatgaacagg
     1321 ggtcagagct cactaaaact tcgagctctc caacagcaga tgagcccacg ataaagattg
     1381 atgatggtcg tgatgagggt aatgaacaag acagctgttc caataccatt aggagaaaaa
     1441 tttccccgtt tgtgatgtca tttggattca gagtatttgg agttgtgctt atcattgtag
     1501 acatcatagt ggtgattgtg gatctggcca tcagtgagaa gaaaagaggc attagagaga
     1561 ttcttgaagg tgtttccctg gctatagcac tcttcttcct tgttgatgtt ctcatgagag
     1621 tgtttgttga aggcttcaag aactatttcc ggtccaaact gaatactttg gatgcagtca
     1681 tagtagtggg cactctgcta attaatatga cctactcctt ctctgacggt acatttcgcg
     1741 gtgataacgt gtatattacc gcggataaac agaaaaacgg cattaaagcg aactttaccg
     1801 tgcgccataa cgtggaagat ggcagcgtgc agctggcgga tcattatcag cagaacaccc
     1861 cgattggcga tggcccggtg ctgctgccgg ataaccatta tctgagcacc cagaccgtgc
     1921 tgagcaaaga tccgaacgaa aaacgcgatc acatggtgct gctggaattt gtgaccgcag
     1981 cgggcattac acacggcatg gatgaactgt atggcggcac cggcggcagc gcgagccagg
     2041 gcgaagaact gtttaccggc gtggtgccga ttctggtgga actggatggc gatgtgaacg
     2101 gccataaatt tagcgtgcgc ggcgaaggcg aaggcgatgc gaccattggc aaactgaccc
     2161 tgaaatttat ttgcaccacc ggcaaactac cggtgccgtg gccgaccctg gtgaccacct
     2221 taacctatgg cgtgcagtgc tttagccgct atccggatca tatgaaacgc catgattttt
     2281 ttaaaagcgc gatgccggaa ggctatgtgc aggaacgcac cattagcttt aaagatgatg
     2341 gcaaatataa aacccgcgcg gtggtgaaat ttgaaggcga taccctggtg aaccgcattg
     2401 aactgaaagg caccgatttt aaagaagatg gcaacattct ggggcataaa ctggaatata
     2461 acacagatca gatgccgcag atggttactc ttcttcgagt tctgagaatt gttatcttaa
     2521 taagaatatt tcagctggct tcacagaaga aacaacttga agtggtaacc ggatcatccg
     2581 gatcttcagg atcctcaatg gtgagcaagg gcgaggagct gatcaaggag aacatgagaa
     2641 gcaagctgta cctggaaggc agcgtgaacg gccaccagtt caagtgcacc cacgaagggg
     2701 agggcaagcc ctacgagggc aagcagaccg cgaggatcaa ggtggtggag ggaggccccc
     2761 tgccgttcgc attcgacatc ctggccacca tgtttatgta cgggagcaag gtgttcatca
     2821 agtaccccgc cgacctcccc gattatttta agcagtcctt ccctgagggc ttcacatggg
     2881 agagagtcat ggtgttcgaa gacgggggcg tgctgaccgc cacccaggac accagcctcc
     2941 aggacggcga gctcatctac aacgtcaagc tcagaggggt gaacttccca gccaacggcc
     3001 ccgtgatgca gaagaaaaca ctgggctggg agcccagcac cgagaccatg acccccgctg
     3061 acggcggcct ggaaggcaga tgcgacaagg tgctgaagct cgtgggcggg ggccacctgc
     3121 acgtcaactt caagaccaca tacaagtcca agaaacccgt gaagatgccc ggcgtccact
     3181 acgtggaccg cagactggaa agaatcaagg aggccgacaa cgagacctac gtcgagcagt
     3241 acgagcacgc tgtggccaga tactccaacc tgggcggagg catggacgag ctgtacaagt
     3301 gataaagctt ggtaccgagc tcggatccac tagtccagtg tggtggaatt ctgcagatat
     3361 ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3421 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3481 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3541 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3601 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3661 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3721 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3781 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3841 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3901 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3961 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4021 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4081 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4141 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4201 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4261 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4321 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4381 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4441 ttggaggcct aggcttttgc aaaaagctcc cgcatgaccg agtacaagcc cacggtgcgc
     4501 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     4561 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     4621 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     4681 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     4741 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     4801 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     4861 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     4921 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     4981 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5041 acctggtgca tgacccgcaa gcccggtgcc tgattcgaaa tgaccgacca agcgacgccc
     5101 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5161 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5221 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5281 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5341 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5401 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5461 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5521 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5581 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5641 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5701 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5761 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5821 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5881 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5941 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6001 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6061 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6121 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6181 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6241 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6301 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6361 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6421 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6481 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6541 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6601 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6661 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6721 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6781 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6841 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6901 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6961 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7021 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7081 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7141 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7201 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7261 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7321 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7381 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7441 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7501 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7561 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     7621 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     7681 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     7741 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     7801 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     7861 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     7921 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     7981 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     8041 tgggggcggg gggggggggg ggggggcgcg sgsggggggg gggggggggg gggggggggg
     8101 ggggggggg
//
