LOCUS       sequence-56515-e        7332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7332)
  AUTHORS   Stutz A, Horvath GL, Monks BG, Latz E
  TITLE     ASC speck formation as a readout for inflammasome activation.
  JOURNAL   Methods Mol Biol. 2013;1040:91-101. doi: 
            10.1007/978-1-62703-523-1_8.
  PUBMED    23852599
REFERENCE   2  (bases 1 to 7332)
  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: "Methods Mol
            Biol."; date: "2013"; volume: "1040:91-101. doi"; pages: " 
            10.1007/978-1-62703-523-1_8"
COMMENT     Sequence Label: pRP_LmCerulean
FEATURES             Location/Qualifiers
     source          1..7332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        215..594
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        595..798
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             799..974
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    1037..1236
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1437..1853
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1783..1805
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1821..1837
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1860..2459
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1860..1879)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2356..2376
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     enhancer        2599..2978
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2979..3182
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3179..3203
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(3365..3367,3368..3370,3371..4084)
                     /codon_start=1
                     /product="monomeric super enhanced variant of CFP (Matsuda 
                     et al., 2008)"
                     /label=mseCFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNDISDNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3410..3431)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3671..3690)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4018..4039
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4138..4731
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4814..4834)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4832..5129
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4999..5134
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5061..5080
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(5175..5191)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5175..5191)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5188..5210)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5199..5215
                     /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(5223..5229,5230..5247,5248..5253))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5268..5289
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5406..5423)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5577..6165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5657..5676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6336..7127,7128..7196))
                     /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     6959..6978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7197..7301)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg agttgacatt gattattgac tagttattaa
      241 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      301 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      361 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      421 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      481 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      541 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      601 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      661 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      721 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      781 gtctatataa gcagagctca ataaaagagc ccacaacccc tcactcggcg cgccagtctt
      841 ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc
      901 cgaatcgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccacgac
      961 gggggtcttt catttggggg ctcgtccggg atttggagac ccctgcccag ggaccaccga
     1021 cccaccaccg ggaggtaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt
     1081 ctatgtttga tgttatgcgc ctgcgtctgt actagttagc taactagctc tgtatctggc
     1141 ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca
     1201 gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga
     1261 ccccgtcagg atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt
     1321 ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca
     1381 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
     1441 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     1501 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     1561 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     1621 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     1681 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     1741 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     1801 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gcctccggta
     1861 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     1921 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     1981 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     2041 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     2101 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     2161 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     2221 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     2281 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     2341 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     2401 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag
     2461 gatcgatccg gccattagcc atattattca ttggttatat agcataaatc aatattggct
     2521 attggccatt gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc
     2581 caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     2641 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     2701 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     2761 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     2821 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     2881 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     2941 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     3001 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     3061 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     3121 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     3181 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     3241 gaagacaccg ggaccgatcc agcctccgcg gccccaagct tccgcgtacg cccgaattcc
     3301 aggatccggg ccgctcgaga gtacagtacc aagggctcgt gaccctccag tcgctaccat
     3361 cgagatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     3421 ggacggcgac gtaaacggcc acaggttcag cgtgtccggc gagggcgagg gcgatgccac
     3481 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     3541 caccctcgtg accaccctga cctggggcgt gcagtgcttc gcccgctacc ccgaccacat
     3601 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgtaccat
     3661 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     3721 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3781 gcacaagctg gagtacaacg acatcagcga caacgtctat atcaccgccg acaagcagaa
     3841 gaacggcatc aaggcccact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3901 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3961 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     4021 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     4081 gtaagcggcc gcatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     4141 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     4201 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     4261 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4321 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     4381 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     4441 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     4501 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     4561 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     4621 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     4681 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc
     4741 gtccgggatc gggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg
     4801 gaactaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4861 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4921 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4981 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5041 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     5101 ttccagaagt agtgaggagg cttttttgga ggcctagtac cgtcgacctc tagctagagc
     5161 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5221 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5281 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5341 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5401 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5461 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5521 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5581 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5641 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5701 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5761 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5821 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5881 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5941 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6001 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6061 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6121 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6181 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6241 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6301 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6361 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6421 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6481 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6541 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6601 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6661 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6721 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6781 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6841 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6901 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6961 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7021 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7081 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7141 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7201 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7261 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7321 ccacctgacg tc
//
