LOCUS       sequence-57154-e        6577 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 6577)
  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 6577)
  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
FEATURES             Location/Qualifiers
     source          1..6577
                     /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"
     LTR             3383..3976
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4059..4079)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4077..4374
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4244..4379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4306..4325
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4420..4436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4420..4436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4433..4455)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4444..4460
                     /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(4468..4474,4475..4492,4493..4498))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4513..4534
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4651..4668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4822..5410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4902..4921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5581..6372,6373..6441))
                     /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     6204..6223
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6442..6546)
                     /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 ggctcgagag gacgcgtgca accggtggtg cggccgcatc gataaaataa aagattttat
     3361 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     3421 aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat
     3481 caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     3541 gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga
     3601 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     3661 ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     3721 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     3781 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc
     3841 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     3901 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     3961 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc
     4021 gacccaccac cgggaggtaa gctgggaact aattctgtgg aatgtgtgtc agttagggtg
     4081 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4141 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4201 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4261 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4321 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4381 agtaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4441 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4501 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4561 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4621 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4681 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4741 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4801 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4861 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4921 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4981 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     5041 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5101 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5161 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5221 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5281 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5341 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5401 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5461 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5521 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5581 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5641 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5701 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5761 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5821 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5881 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5941 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6001 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6061 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6121 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6181 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6241 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6301 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6361 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6421 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6481 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6541 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
