LOCUS       sequence-378633-        6675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BMAL1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6675)
  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 6675)
  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-BMAL1
FEATURES             Location/Qualifiers
     source          1..6675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1601..1623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1685..1704
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3696..3715)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3761..3816
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3815..3834)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4177..4193)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4177..4193)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4190..4212)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4201..4217
                     /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(4225..4231,4232..4249,4250..4255))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4270..4291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4350..4545
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4396..4531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4551..4685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4588..4607)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4642..4661
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4753..4770)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4924..5512)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5004..5023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5683..6474,6475..6543))
                     /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     6306..6325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6544..6648)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg ccrggsgggg sggggsgggg sgrggggsgg
      541 ggsggggsgr ggsggrgrgg kgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccggggr gggctcgggg grggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt catggcggac
     1741 cagagaatgg acatttcctc aaccatcagc gacttcatgt ctccgggccc caccgaccta
     1801 ctctccggtt ccctgggcac cagtggtgtg gactgcaatc gcaagaggaa aggcagtgcc
     1861 actgactacc aagaaagtat ggacacagac aaagatgacc ctcatggaag gttagaatat
     1921 gcagaacacc aaggaaggat caagaatgca agggaggccc acagtcagat tgaaaagagg
     1981 cgtcgggaca aaatgaacag tttcattgat gaattggctt ctttggtacc aacatgcaat
     2041 gcaatgtcca ggaagttaga taaactcacc gtgctaagga tggctgttca gcacatgaaa
     2101 actttgagag gtgccaccaa cccatacaca gaagcaaact acaagccaac atttctatca
     2161 gatgacgaac tgaaacacct aattctcagg gcagcagatg gatttttgtt tgtcgtagga
     2221 tgtgaccgag ggaagatcct ctttgtctcc gagtctgtct tcaagatcct caattatagc
     2281 cagaatgacc ttattggcca gagcttgttt gactacctgc atccaaaaga tattgccaaa
     2341 gttaaggaac agctatcttc ctcggacact gcgccccggg agcgactcat tgatgccaag
     2401 actggacttc cggttaaaac ggatataacc cctgggccct cccggctatg ctctggagcc
     2461 cgccgctctt tcttctgtag aatgaagtgc aacaggcctt cagtaaaggt ggaagataag
     2521 gacttcgcct ctacctgttc aaagaaaaaa gcagatcgaa aaagcttctg cacaatccac
     2581 agcacaggct atttgaaaag ctggccaccc acgaagatgg ggctggacga agacaatgag
     2641 ccagacaacg agggctgcaa cctcagctgc ctcgttgcaa tcgggcgcct gcactcgcac
     2701 atggttccac aaccagcgaa cggggaaata cgggtgaaat ctatggagta cgtttctcga
     2761 cacgcaatag atgggaaatt tgtttttgta gatcagaggg cgacagctat tttggcgtat
     2821 ctaccacagg aacttctagg tacatcatgt tatgagtatt ttcatcaaga cgacatagga
     2881 cacctcgcag aatgtcacag gcaagtttta cagacaagag aaaagatcac aactaattgc
     2941 tataagttta agatcaaaga tggttctttt atcacgctac gaagtcgatg gttcagtttc
     3001 atgaacccgt ggaccaagga agttgaatac attgtctcaa ccaacactgt tgttttagcc
     3061 aatgtcctgg aaggcgggga cccaaccttc ccgcagctaa cagcaccccc ccacagcatg
     3121 gacagcatgc tgccctctgg agaaggtggc ccaaagagga ctcatcccac tgtcccaggc
     3181 attccagggg gaaccagagc cggagcagga aaaataggtc gaatgatcgc ggaggaaatc
     3241 atggaaatcc acaggataag agggtcatcg ccttccagct gtggctccag cccgctgaac
     3301 atcacaagta cgcctccccc tgatgcctct tctccaggag gcaagaagat tctaaatgga
     3361 gggactccag acattccttc cactggacta ttaccagggc aggctcagga gaccccaggg
     3421 tatccctatt ctgatagttc ttctattctt ggtgagaacc cccacatagg catcgatatg
     3481 atagataacg accaaggatc aagtagtccc agtaacgatg aggcagcaat ggctgtcatc
     3541 atgagcctct tggaagcaga tgcggggctg ggtggccccg ttgactttag tgacttgcca
     3601 tggccgctgt aggaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     3661 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     3721 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     3781 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     3841 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     3901 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     3961 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     4021 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     4081 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     4141 tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4201 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4261 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4321 gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg
     4381 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4441 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4501 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     4561 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4621 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4681 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4741 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4801 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4861 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4921 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4981 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5041 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5101 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5161 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5221 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5281 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5341 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5401 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5461 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5521 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5581 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5641 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5701 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5761 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5821 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5881 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5941 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6001 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6061 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6121 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6181 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6241 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6301 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6361 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6421 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6481 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6541 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6601 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6661 aaaagtgcca cctgg
//
