LOCUS       sequence-78756-e        5702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Flag-tagged mAIM2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5702)
  AUTHORS   Fernandes-Alnemri T, Yu JW, Datta P, Wu J, Alnemri ES
  TITLE     AIM2 activates the inflammasome and cell death in response to 
            cytoplasmic DNA.
  JOURNAL   Nature. 2009 Mar 26;458(7237):509-13. doi: 10.1038/nature07710. Epub
            2009 Jan 21.
  PUBMED    19158676
REFERENCE   2  (bases 1 to 5702)
  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"; doi: 
            "10.1038/nature07710"; journalName: "Nature"; date: "2009-03-26- 
            26"; volume: "458"; issue: "7237"; pages: "509-13"
COMMENT     Sequence Label: pFlag-CMV2-mAIM2
FEATURES             Location/Qualifiers
     source          1..5702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2043..2665
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2242..2261)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2689..2709)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2694..3023
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2874..3009
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3061..3080)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3062..3080)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3094..3110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3094..3110)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3107..3129)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3118..3134
                     /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(3142..3148,3149..3166,3167..3172))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3187..3208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3325..3342)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3496..4084)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3576..3595)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4255..5046,5047..5115))
                     /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     4878..4897
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5116..5220)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5247..5675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5334..5353)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5544..5565
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tgagagtgag taccgggaaa tgctgttgtt gaccggcctg gaccacatca cggaggaaga
     1021 actgaaacgg ttcaagtact ttgctttgac tgagtttcag attgccagga gcacactcga
     1081 cgtggcagat aggacagagt tagctgacca cctgattcaa agtgcaggtg cggcgtctgc
     1141 agtgaccaag gccattaata ttttccagaa gttgaattat atgcatattg caaatgctct
     1201 tgaagagaaa aagaaagaag ctgaacgtaa actcatgacc aatacaaaga agagaggaac
     1261 acagaaggta gaaaatagaa gtcaagctga aaactgctct gctgcctctg ccacccgcag
     1321 tgacaatgac tttaaggaac aggctgctac agaagtctgt cctcaagcta agcctcagaa
     1381 gaaacagatg gtggcagaac aggaagccat cagagaagat ttacagaaag atccacttgt
     1441 tgtcacggtg ctgaaagcta taaatccctt tgagtgtgag actcaggaag gaagacaaga
     1501 gatatttcat gcaacagtgg ccacggagac agattttttc tttgtaaaag ttttaaacgc
     1561 acagtttaaa gataaattta tcccaaagag gacaattaaa atatcaaact acctttggca
     1621 cagtaacttc atggaggtca ccagttcctc agttgtggtt gatgttgaat ctaaccacga
     1681 agtcccaaat aacgttgtta agagagccag ggaaactccc aggattagta aactgaagat
     1741 tcagccatgt ggaacaattg tgaatgggct gtttaaagtc cagaagataa cagaggaaaa
     1801 agatagagta ctgtatggta tacatgataa aacagggaca atggaggtgt tggtgctggg
     1861 aaacccaagc aaaacaaagt gcgaggaagg agacaagatt agactcacgt tctttgaggt
     1921 gtcaaaaaat ggagtgaaaa ttcagttgaa atctggacct tgtagctttt ttaaggttat
     1981 taaggctgca aagccaaaaa ctgacatgaa aagtgtggag tgactcgact ctagaggatc
     2041 ccgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc
     2101 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt
     2161 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga
     2221 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt
     2281 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt
     2341 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac
     2401 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac
     2461 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc
     2521 tgattttaaa ataactatac cagcaggagg acgtccagac acagcatagg ctacctggcc
     2581 atgcccaacc ggtgggacat ttgagttgct tgcttggcac tgtcctctca tgcgttgggt
     2641 ccactcagta gatgcctgtt gaattgggta cgcggccagc ttggctgtgg aatgtgtgtc
     2701 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2761 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2821 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2881 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2941 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3001 ttggaggcct aggcttttgc aaaaagctcc tcgaggaact gaaaaaccag aaagttaatt
     3061 ccctatagtg agtcgtatta aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     3121 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3181 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3241 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3301 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3361 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3421 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3481 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3541 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3601 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3661 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3721 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3781 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3841 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3901 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3961 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4021 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4081 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4141 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4201 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4261 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4321 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4381 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4441 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4501 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4561 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4621 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4681 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4741 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4801 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4861 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4921 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4981 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5041 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5101 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5161 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5221 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     5281 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5341 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5401 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5461 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5521 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5581 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5641 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     5701 tt
//
