LOCUS       sequence-261281-        7478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET-based sensor for CaMKII activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7478)
  AUTHORS   Ardestani G, West MC, Maresca TJ, Fissore RA, Stratton MM
  TITLE     FRET-based sensor for CaMKII activity (FRESCA): A useful tool for 
            assessing CaMKII activity in response to Ca(2+) oscillations in live
            cells.
  JOURNAL   J Biol Chem. 2019 Jun 14. pii: RA119.009235. doi: 
            10.1074/jbc.RA119.009235.
  PUBMED    31201271
REFERENCE   2  (bases 1 to 7478)
  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: "J Biol 
            Chem. 2019 Jun 14. pii: RA119.009235. doi: 
            10.1074/jbc.RA119.009235."
COMMENT     Sequence Label: FRESCA-syntide
FEATURES             Location/Qualifiers
     source          1..7478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(923..925,926..928,929..1639)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2204..2206,2207..2209,2210..2923)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2936..2977
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2987..3004
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3069..3086)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3075..3299
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3345..3773
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3432..3451)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3642..3663
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3782..3802)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3787..4116
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3967..4102
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4029..4048
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4183..4977
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4237..4256)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4847..4866
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5151..5272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5188..5207)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5242..5261
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5321..5337)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5321..5337)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5334..5356)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5345..5361
                     /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(5369..5375,5376..5393,5394..5399))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5414..5435
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5552..5569)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5723..6311)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5803..5822)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6482..7273,7274..7342))
                     /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     7105..7124
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7343..7447)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccatggtgag caagggcgag gagctgttca ccggggtggt
      961 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     1021 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1081 gctgcccgtg ccctggccca ccctcgtgac caccctgtcc tggggcgtgc agtgcttcgc
     1141 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1201 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1261 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1321 ggacggcaac atcctggggc acaagctgga gtacaactac tttagcgaca acgtctatat
     1381 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
     1441 ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1501 cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa
     1561 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1621 catggacgag ctgtacaaga ctagtaaggg taatggtagg tttttaactt taaaaccatt
     1681 gcctgacagc attattcaag aaagcctgga gattcagcaa ggtgtgaatc catttttcat
     1741 tggtagatcc gaggattgca attgtaaaat tgaagacagt aggttgtctc gagttcattg
     1801 cttcattttc aaaaagaggc atgctgtagg caaaagcatg tatgaatctc cggcacaagg
     1861 tttagatgat atttggtatt gccacaccgg aactaacgtg agctatttaa ataataaccg
     1921 catgatacag ggtacgaaat tccttttaca agacggagat gaaatcaaga tcatttggga
     1981 taaaaacaat aaatttgtca ttggctttaa agtggaaatt aacgatacta caggtctgtt
     2041 taacgaggga ttaggtatgt tacaagaaca aagagtagta cttaagcaaa cagccgaaga
     2101 aaaagatttg gtgaaaaagt taggaggatc cggcggcgcc ggcggcggcc cgctggcgcg
     2161 cgccctgacc gtggcgggcc tgccgggcaa aaaaggcggc cgcatggtga gcaagggcga
     2221 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2281 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2341 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg
     2401 ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2461 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2521 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2581 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2641 caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     2701 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa
     2761 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     2821 caaactgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     2881 cgccgccggg atcactctcg gcatggacga gctgtacaag taaccgcggt tcgaaggtaa
     2941 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca
     3001 ccattgagtt tatccagcac agtggcggcc gctcgagtct agagggcccg tttaaacccg
     3061 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3121 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3181 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3241 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3301 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3361 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3421 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3481 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3541 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3601 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3661 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3721 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3781 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3841 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3961 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4021 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4081 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4141 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4201 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4261 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4321 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4381 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4441 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4501 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4561 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4621 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4681 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4741 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4801 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4861 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4921 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4981 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5041 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5101 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5161 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5221 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5281 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5341 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5401 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5461 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5521 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5581 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5641 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5701 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5761 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5821 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5881 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5941 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6001 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6061 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6121 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6181 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6241 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6301 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6361 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6421 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6481 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6541 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6601 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6661 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6721 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6781 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6841 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6901 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6961 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7021 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7081 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7141 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7201 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7261 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7321 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7381 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7441 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
