LOCUS       sequence-370058-        6336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-CaMKIIG in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6336)
  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 6336)
  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-CaMKIIG
FEATURES             Location/Qualifiers
     source          1..6336
                     /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(3358..3377)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3423..3478
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3477..3496)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3839..3855)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3839..3855)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3852..3874)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3863..3879
                     /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(3887..3893,3894..3911,3912..3917))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3932..3953
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4012..4207
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4058..4193
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4213..4347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4250..4269)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4304..4323
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4415..4432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4586..5174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4666..4685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5345..6136,6137..6205))
                     /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     5968..5987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6206..6310)
                     /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 rggcggagag 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 cgccaccacc gccacctgca cccgcttcac
     1801 cgacgactac cagcttttcg aggagctcgg caagggtgcc ttctcagtgg tccggaggtg
     1861 tgtaaagaaa acatctacgc aggaatatgc tgcaaaaatc atcaatacca agaagttgtc
     1921 cgcccgagat catcagaaac tagaacgaga ggcccggata tgccgacttc tgaaacatcc
     1981 aaacattgtg cgtctccatg acagtatttc tgaagaaggg tttcactacc tcgtgtttga
     2041 ccttgttacc ggaggggagt tgtttgaaga cattgtggcc agagagtact acagtgaagc
     2101 tgatgccagc cactgtatac atcagatcct ggagagtgtt aaccacatcc accagcatga
     2161 catcgtccac cgggatctga agcctgagaa cttgctgctg gcgagtaaat gcaagggtgc
     2221 tgcggtcaag ctagctgatt ttggcctggc catcgaagta cagggagagc agcaggcttg
     2281 gtttggtttt gccggcaccc caggttactt gtcccctgag gtcttgagga aagatcccta
     2341 tggaaaacct gtggacatct gggcctgcgg ggtcatcctg tatatcctcc tggtgggcta
     2401 ccctcccttc tgggatgagg atcagcacaa gctgtatcag cagatcaaag ctggagccta
     2461 cgatttccca tcaccagaat gggacacagt cactcctgaa gctaagaact tgatcaacca
     2521 gatgctgacc ataaaccctg caaagcgcat cacggccgac caggctctca agcacccatg
     2581 ggtctgtcaa cggtctacgg tggcatccat gatgcatcgc caagagacgg tagagtgctt
     2641 acgcaaattc aacgcccgga gaaaactgaa gggtgccatc ctcacaacca tgcttgtctc
     2701 caggaacttt tcagctgcca aaagcctatt gaacaagaag tcagatggcg gtgtcaagga
     2761 accacaaacc accgtggtac ataatgctac agatgggatc aagggctcca cagagagctg
     2821 taacaccact acagaagacg aagatctcaa agtgagaaaa caagaaatca ttaagatcac
     2881 agaacaactg atcgaagcca tcaacaatgg ggactttgag gcctacacga agatttgtga
     2941 cccgggcctc acatcctttg agccagaagc ccttggtaac ctcgtggaag gaatggattt
     3001 ccataagttt tactttgaga atctcctgtc caagaacagc aagcctatcc acaccaccat
     3061 cctaaaccct cacgtccacg tgattgggga ggacgcagct tgcatcgcct atatccggct
     3121 cactcagtac atcgacgggc agggtcggcc tcgcaccagt cagtcagagg agacccgggt
     3181 gtggcaccgc cgggatggca agtggctcaa tgtccactat cactgctcag gggcccctgc
     3241 cgcaccgcta cagtgaacgc gtgaaggtac cagggaattc actcctcagg tgcaggctgc
     3301 ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct
     3361 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     3421 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     3481 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     3541 tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca
     3601 tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg
     3661 aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt
     3721 ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc
     3781 tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt
     3841 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3901 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3961 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa
     4021 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4081 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4141 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4201 ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4261 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4321 atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga
     4381 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4441 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4501 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4561 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4621 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4681 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4741 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4801 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4861 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4921 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4981 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5041 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5101 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5161 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5221 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5281 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5341 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5401 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5461 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5521 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5581 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5641 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5701 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5761 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5821 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5881 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5941 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6001 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6061 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6121 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6181 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6241 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6301 gggttccgcg cacatttccc cgaaaagtgc cacctg
//
