LOCUS       sequence-379366-        6285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-CaMKIID-K43M in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6285)
  AUTHORS   Li H, Ou J, Li Y, Xu N, Li Q, Wu P, Peng C, Tang YC, Chang HC
  TITLE     ISX-9 potentiates CaMKIIdelta-mediated BMAL1 activation to enhance 
            circadian amplitude.
  JOURNAL   Commun Biol. 2022 Jul 28;5(1):750. doi: 10.1038/s42003-022-03725-x.
  PUBMED    35902736
REFERENCE   2  (bases 1 to 6285)
  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: "Commun 
            Biol."; date: "2022-07-28"; volume: "5(1):750. doi"; pages: " 
            10.1038/s42003-022-03725-x"
COMMENT     Sequence Label: pCAGGS-FLAG-CaMKIID-K43M
FEATURES             Location/Qualifiers
     source          1..6285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1739..1762
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3307..3326)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3372..3427
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3426..3445)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3788..3804)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3788..3804)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3801..3823)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3812..3828
                     /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(3836..3842,3843..3860,3861..3866))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3881..3902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3961..4156
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4007..4142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4162..4296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4199..4218)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4253..4272
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4364..4381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4535..5123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4615..4634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5294..6085,6086..6154))
                     /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     5917..5936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6155..6259)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccrggsggg gsggggsggg gsgrggggsg
      541 gggsggggsg rggsggrgag gkgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg rgggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcaccatgga
     1741 ttacaaggat gacgacgata agtgcggccg cgcttcgacc accacctgca cccggttcac
     1801 cgacgagtat cagctctttg aggagctcgg aaagggggcg ttctcagtgg tgagaagatg
     1861 tatgaaaatc cctactggac aagagtatgc tgccatgatt atcaacacca aaaagctttc
     1921 tgctagggac catcagaaac tggaaaggga agctagaatc tgccgtctct tgaagcaccc
     1981 caatattgtg agacttcacg acagtatatc ggaggagggc ttccattact tggtgtttga
     2041 cttagtgact ggtggcgaac tgtttgaaga catagtggca agagaatatt acagtgaagc
     2101 tgatgccagt cattgtatac aacagattct agagagtgta aatcattgtc acctaaatgg
     2161 catagttcac agggacctga agcctgagaa tttgctttta gctagcaagt ccaaaggagc
     2221 agctgtgaag ctggcagact tcggcttagc catagaagtt caaggcgacc agcaggcatg
     2281 gtttggtttt gctggcacac ctgggtatct ttctccagaa gtcctgcgta aagatcctta
     2341 tggaaaacca gtggatatgt gggcatgcgg tgtcatcctc tacatcttgc tggtgggata
     2401 cccacccttc tgggatgaag atcagcatag actgtatcag cagatcaagg ccggagctta
     2461 cgattttccg tcaccagaat gggatacagt gacacctgaa gccaaagacc tcatcaacaa
     2521 aatgctgacc atcaaccctg ccaaacgtat cacagcctct gaggccctga aacacccatg
     2581 gatctgtcaa cgctctactg ttgcctccat gatgcacagg caggagactg tagactgctt
     2641 gaagaaattt aatgctagac ggaaactgaa gggcgccatc ttgacaacta tgctggctac
     2701 gagaaatttt tcagcagcca agagtttatt gaagaaacca gatggggtaa aggagtcaac
     2761 tgagagctca aacaccacca ttgaggatga agacgtgaaa gcacgaaaac aggagatcat
     2821 caaagtcact gagcaactga ttgaagctat caacaatggg gactttgagg cttacacaaa
     2881 aatctgtgat ccaggcctca ctgcctttga acctgaagca ttgggcaact tagtggaagg
     2941 gatggacttt cacagattct actttgaaaa tgctttgtcc aaaagcaata aaccaatcca
     3001 cacgatcatc ctcaacccac acgttcacct ggtaggggat gacgcagcct gcatcgcata
     3061 cattcggctc acacagtaca tggacggaag cgggatgcca aagaccatgc agtcagaaga
     3121 gacgcgcgtg tggcaccgcc gtgatgggaa gtggcagaat gttcactttc accgttcggg
     3181 gtcccccaca gtacccatca actaaacgcg tgaaggtacc agggaattca ctcctcaggt
     3241 gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca
     3301 ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     3361 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     3421 tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt
     3481 ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata
     3541 aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc
     3601 cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc
     3661 cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc
     3721 agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt
     3781 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3841 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3901 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc
     3961 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4021 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4081 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4141 cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc
     4201 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4261 tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac
     4321 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4381 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4441 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4501 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4561 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4621 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4681 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4741 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4801 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4861 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4921 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4981 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5041 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5101 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5161 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5221 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5281 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5341 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5401 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5461 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5521 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5581 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5641 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5701 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5761 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5821 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5881 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5941 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6001 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6061 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6121 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6181 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6241 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctg
//
