LOCUS       sequence-370052-        6730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-BMAL1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6730)
  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 6730)
  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-BMAL1
FEATURES             Location/Qualifiers
     source          1..6730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1740..1763
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3752..3771)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3817..3872
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3871..3890)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4233..4249)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4233..4249)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4246..4268)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4257..4273
                     /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(4281..4287,4288..4305,4306..4311))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4326..4347
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4406..4601
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4452..4587
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4607..4741
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4644..4663)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4698..4717
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4809..4826)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4980..5568)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5060..5079)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5739..6530,6531..6599))
                     /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     6362..6381
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6600..6704)
                     /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 gggggnsgcg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggs gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg grgggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcaccatgg
     1741 attacaagga tgacgacgat aagtgcggcc gcgcggacca gagaatggac atttcctcaa
     1801 ccatcagcga cttcatgtct ccgggcccca ccgacctact ctccggttcc ctgggcacca
     1861 gtggtgtgga ctgcaatcgc aagaggaaag gcagtgccac tgactaccaa gaaagtatgg
     1921 acacagacaa agatgaccct catggaaggt tagaatatgc agaacaccaa ggaaggatca
     1981 agaatgcaag ggaggcccac agtcagattg aaaagaggcg tcgggacaaa atgaacagtt
     2041 tcattgatga attggcttct ttggtaccaa catgcaatgc aatgtccagg aagttagata
     2101 aactcaccgt gctaaggatg gctgttcagc acatgaaaac tttgagaggt gccaccaacc
     2161 catacacaga agcaaactac aagccaacat ttctatcaga tgacgaactg aaacacctaa
     2221 ttctcagggc agcagatgga tttttgtttg tcgtaggatg tgaccgaggg aagatcctct
     2281 ttgtctccga gtctgtcttc aagatcctca attatagcca gaatgacctt attggccaga
     2341 gcttgtttga ctacctgcat ccaaaagata ttgccaaagt taaggaacag ctatcttcct
     2401 cggacactgc gccccgggag cgactcattg atgccaagac tggacttccg gttaaaacgg
     2461 atataacccc tgggccctcc cggctatgct ctggagcccg ccgctctttc ttctgtagaa
     2521 tgaagtgcaa caggccttca gtaaaggtgg aagataagga cttcgcctct acctgttcaa
     2581 agaaaaaagc agatcgaaaa agcttctgca caatccacag cacaggctat ttgaaaagct
     2641 ggccacccac gaagatgggg ctggacgaag acaatgagcc agacaacgag ggctgcaacc
     2701 tcagctgcct cgttgcaatc gggcgcctgc actcgcacat ggttccacaa ccagcgaacg
     2761 gggaaatacg ggtgaaatct atggagtacg tttctcgaca cgcaatagat gggaaatttg
     2821 tttttgtaga tcagagggcg acagctattt tggcgtatct accacaggaa cttctaggta
     2881 catcatgtta tgagtatttt catcaagacg acataggaca cctcgcagaa tgtcacaggc
     2941 aagttttaca gacaagagaa aagatcacaa ctaattgcta taagtttaag atcaaagatg
     3001 gttcttttat cacgctacga agtcgatggt tcagtttcat gaacccgtgg accaaggaag
     3061 ttgaatacat tgtctcaacc aacactgttg ttttagccaa tgtcctggaa ggcggggacc
     3121 caaccttccc gcagctaaca gcaccccccc acagcatgga cagcatgctg ccctctggag
     3181 aaggtggccc aaagaggact catcccactg tcccaggcat tccaggggga accagagccg
     3241 gagcaggaaa aataggtcga atgatcgcgg aggaaatcat ggaaatccac aggataagag
     3301 ggtcatcgcc ttccagctgt ggctccagcc cgctgaacat cacaagtacg cctccccctg
     3361 atgcctcttc tccaggaggc aagaagattc taaatggagg gactccagac attccttcca
     3421 ctggactatt accagggcag gctcaggaga ccccagggta tccctattct gatagttctt
     3481 ctattcttgg tgagaacccc cacataggca tcgatatgat agataacgac caaggatcaa
     3541 gtagtcccag taacgatgag gcagcaatgg ctgtcatcat gagcctcttg gaagcagatg
     3601 cggggctggg tggccccgtt gactttagtg acttgccatg gccgctgtag acgcgtgaag
     3661 gtaccaggga attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     3721 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     3781 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     3841 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     3901 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     3961 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     4021 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     4081 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     4141 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     4201 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4261 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4321 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4381 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc
     4441 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4501 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4561 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
     4621 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4681 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     4741 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4801 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4861 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4921 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4981 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5041 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5101 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5161 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5221 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5281 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5341 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5401 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5461 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5521 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5581 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5641 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5701 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5761 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5821 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5881 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5941 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6001 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6061 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6121 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6181 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6241 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6301 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6361 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6421 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6481 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6541 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6601 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6661 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6721 gtgccacctg
//
