LOCUS       sequence-272044-        5760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mCyRFP2-CREB donor for R-CREB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5760)
  AUTHORS   Laviv T, Scholl B, Parra-Bueno P, Foote B, Zhang C, Yan L, Hayano Y,
            Chu J, Yasuda R
  TITLE     In Vivo Imaging of the Coupling between Neuronal and CREB Activity 
            in the Mouse Brain.
  JOURNAL   Neuron. 2019 Dec 17. pii: S0896-6273(19)31037-2. doi: 
            10.1016/j.neuron.2019.11.028.
  PUBMED    31883788
REFERENCE   2  (bases 1 to 5760)
  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: "Neuron. 
            2019 Dec 17. pii: S0896-6273(19)31037-2. doi: 
            10.1016/j.neuron.2019.11.028."
COMMENT     Sequence Label: CMV-mCyRFP2-CREB
FEATURES             Location/Qualifiers
     source          1..5760
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1314
                     /codon_start=1
                     /product="monomeric cyan-excitable red fluorescent protein 
                     (Laviv et al., 2016)"
                     /label=mCyRFP1
                     /translation="MVSKGEELIKEEMTSKLYLEGSVNGHYFKCTHEGEGKPYEGKQTA
                     RIKVVEGGPLPFAFDILATHFMYGSKVFIKYPADIPDYFKQSFPEGFTWERVMVFEDGG
                     VLTTTQDTSLQDGVLIYNVKTRGVNFPANGPVMQKKTLGWEPSTETMYPADGGLEGRCD
                     KKLKLVGGGHLHVNFKTTYKSKKPVKMPGVHYVDRRLERIKEADNETYVEQYEHAVARY
                     SNLGGGMDELYK"
     misc_feature    2359..2424
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2548..2669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2585..2604)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2639..2658
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2676..3131)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2813..2834)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3025..3044
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3158..3262
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3264..3621
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3264..3455
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3287..3307)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3472..3607
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3656..4450
                     /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             3656..4450
                     /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(3710..3729)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4320..4339
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4638..4657)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4682..4729
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5058..5646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5547..5566
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgatca aggaggaaat gaccagcaag
      661 ctgtacctgg aaggcagcgt gaacggccac tacttcaagt gcacccacga aggggagggc
      721 aagccctacg agggcaagca gaccgccagg atcaaggtgg tggagggagg ccccctgccg
      781 ttcgcattcg acatcctggc cacccacttt atgtacggga gcaaggtgtt catcaagtac
      841 cccgccgaca tccccgatta ttttaagcag tccttccctg agggcttcac atgggagaga
      901 gtcatggtgt tcgaagacgg gggcgtgctg accactaccc aggacaccag cctccaggac
      961 ggcgtgctca tctacaacgt caagaccaga ggggtgaact tcccagccaa cggccccgtg
     1021 atgcagaaga aaacactggg ctgggagccc agcaccgaga ccatgtaccc cgccgacggc
     1081 ggcctggaag gcagatgcga caagaagctg aagctcgtgg gcgggggcca cctgcacgtc
     1141 aacttcaaga ccacatacaa gtccaagaaa cccgtgaaga tgcccggcgt ccactacgtg
     1201 gacaggagac tggaaagaat caaggaggcc gacaacgaga cctacgtcga gcagtacgag
     1261 cacgctgtgg ccagatactc caacctgggc ggaggcatgg acgagctgta caagcctacc
     1321 cctaccccta ccatgaccat ggactctgga gcagacaacc agcagagtgg agatgctgct
     1381 gtaacagaag ctgaaagtca acaaatgaca gttcaagccc agccacagat tgccacatta
     1441 gcccaggtat ccatgccagc agctcatgcg acgtcatctg ctcccactgt aaccttagtg
     1501 cagctgccca atgggcagac agtccaggtc catggggtca tccaggcggc ccagccatca
     1561 gttattcagt ctccacaagt ccaaacagtt cagtcttcct gtaaggactt aaaaagactt
     1621 ttctccggaa ctcagatttc aactattgca gaaagtgaag attcacagga gtctgtggat
     1681 agtgtaactg attcccaaaa acgaagggaa atcctttcaa ggaggccttc ctacaggaaa
     1741 attttgaatg acttatcttc tgatgcacca ggggtgccaa ggattgaaga agaaaaatca
     1801 gaagaagaga cttcagcccc tgccatcacc actgtaacag tgccaacccc gatttaccaa
     1861 actagcagtg ggcagtatat tgccattacc cagggaggag caatacagct ggctaacaat
     1921 ggtaccgatg gggtacaggg cctgcagaca ttaaccatga ccaatgcagc tgccactcag
     1981 ccgggtacta ccattctaca atatgcacag accactgatg gacagcagat tctagtgccc
     2041 agcaaccaag ttgttgttca agctgcctct ggtgatgtac aaacatacca gattcgcaca
     2101 gcacccacta gcaccattgc ccctggagtt gttatggcgt cctccccagc acttcctaca
     2161 cagcctgctg aagaagcagc acgaaagaga gaggttcgtc taatgaagaa cagggaagca
     2221 gcaagagaat gtcgtagaaa gaagaaagaa tatgtgaaat gtttagagaa cagagtggca
     2281 gtgcttgaaa accaaaacaa aacattgatt gaggagctaa aagcacttaa ggacctttac
     2341 tgccacaagt cagattgatc cggactcaga tctcgagctc aagcttcgaa ttctgcagtc
     2401 gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc
     2461 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2521 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2581 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2641 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     2701 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2761 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2821 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2881 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2941 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3001 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3061 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3121 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3181 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3241 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3301 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3361 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3421 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3481 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3541 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3601 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     3661 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3721 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3781 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     3841 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3901 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3961 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4021 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4081 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4141 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4201 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4261 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4321 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4381 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4441 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4501 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4561 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4621 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4681 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     4741 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4801 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4861 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     4921 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4981 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5041 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5101 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5161 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5221 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5281 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5341 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5401 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5461 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5521 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5581 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5641 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5701 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
