LOCUS       sequence-180720-        5430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP tagged zebrafish ASC in the pCS2+ vector for in vitro 
            transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5430)
  AUTHORS   Vincent WJ, Freisinger CM, Lam PY, Huttenlocher A, Sauer JD
  TITLE     Macrophages mediate flagellin induced inflammasome activation and 
            host defense in zebrafish.
  JOURNAL   Cell Microbiol. 2016 Apr;18(4):591-604. doi: 10.1111/cmi.12536. Epub
            2015 Nov 4.
  PUBMED    26468080
REFERENCE   2  (bases 1 to 5430)
  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: "10.1111/cmi.12536";
            journalName: "Cell Microbiol"; date: "2016-04"; volume: "18"; issue:
            "4"; pages: "591-604"
COMMENT     Sequence Label: pCS2+ gfp-zASC
FEATURES             Location/Qualifiers
     source          1..5430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      92..101
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             98..814
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    1479..1613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1503..1522)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1557..1576
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1726..1746)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1726..1744)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1765..1781)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1765..1781)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1778..1800)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1789..1805
                     /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(1813..1819,1820..1837,1838..1843))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1858..1879
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1996..2013)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2167..2755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2247..2266)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2926..3717,3718..3786))
                     /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     3549..3568
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3787..3891)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3917..4372)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4054..4075)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4266..4285
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4446..5430
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccaccggt cgccaccatg gtgagcaagg gcgccgagct
      121 gttcaccggc atcgtgccca tcctgatcga gctgaatggc gatgtgaatg gccacaagtt
      181 cagcgtgagc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
      241 ctgcaccacc ggcaagctgc ctgtgccctg gcccaccctg gtgaccaccc tgagctacgg
      301 cgtgcagtgc ttctcacgct accccgatca catgaagcag cacgacttct tcaagagcgc
      361 catgcctgag ggctacatcc aggagcgcac catcttcttc gaggatgacg gcaactacaa
      421 gtcgcgcgcc gaggtgaagt tcgagggcga taccctggtg aatcgcatcg agctgaccgg
      481 caccgatttc aaggaggatg gcaacatcct gggcaataag atggagtaca actacaacgc
      541 ccacaatgtg tacatcatga ccgacaaggc caagaatggc atcaaggtga acttcaagat
      601 ccgccacaac atcgaggatg gcagcgtgca gctggccgac cactaccagc agaatacccc
      661 catcggcgat ggccctgtgc tgctgcccga taaccactac ctgtccaccc agagcgccct
      721 gtccaaggac cccaacgaga agcgcgatca catgatctac ttcggcttcg tgaccgccgc
      781 cgccatcacc cacggcatgg atgagctgta caagaattca gatctcaaga tggcggaatc
      841 tttcaaggag cacctgcagg aggcttttga ggatctgggt gctgataatc ttagaaagtt
      901 taaaagcaaa ctgggcgatc ggaggcagga accgcgcgtc acaaagtctg caatcgaaaa
      961 gctgaaagac gagatcgatc tggctgattt aatggtcggc gtgttcacat caaaagacgc
     1021 ggtttctgta acagtggaga ttttgagggc gatcaagtgt attgcagtag cagatgatct
     1081 attgaggaac acagggcaat cagaaagcaa aggtgctccc tctgatgaaa gcaaatgtgc
     1141 ttcctctaag gctgtaagca aagttgcttt ctccaaggta aacttcatcg atgagcactg
     1201 gaaagaattg atagaccgag ttaataatgt ggaccccatc ctagatatac ttcggcagaa
     1261 aaaagtcatc acaaatgagg attactgtac catccgtaat aaggagactc cccaaaagaa
     1321 gatgagagag ttattaacag gcccaatcac atgtgctggc aacaaaggga aagaagtctt
     1381 gtatgatgct ctcagagaga gcaataaatt cctgatggat gaccttgagg atgctcagtg
     1441 actcgagcct ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca
     1501 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1561 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1621 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg
     1681 cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt
     1741 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1801 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1861 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1921 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1981 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2041 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2101 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2161 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2221 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2281 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2341 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2401 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2461 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2521 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2581 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     2641 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2701 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2761 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2821 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2881 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2941 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3001 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3061 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3121 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3181 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3241 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3301 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3361 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3421 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3481 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3541 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3601 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3661 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3721 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3781 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3841 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3901 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3961 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4021 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4081 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4141 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4201 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4261 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4321 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     4381 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4441 cagtcgacca tagccaattc aatatggcgt atatggactc atgccaattc aatatggtgg
     4501 atctggacct gtgccaattc aatatggcgt atatggactc gtgccaattc aatatggtgg
     4561 atctggaccc cagccaattc aatatggcgg acttggcacc atgccaattc aatatggcgg
     4621 acttggcact gtgccaactg gggaggggtc tacttggcac ggtgccaagt ttgaggaggg
     4681 gtcttggccc tgtgccaagt ccgccatatt gaattggcat ggtgccaata atggcggcca
     4741 tattggctat atgccaggat caatatatag gcaatatcca atatggccct atgccaatat
     4801 ggctattggc caggttcaat actatgtatt ggccctatgc catatagtat tccatatatg
     4861 ggttttccta ttgacgtaga tagcccctcc caatgggcgg tcccatatac catatatggg
     4921 gcttcctaat accgcccata gccactcccc cattgacgtc aatggtctct atatatggtc
     4981 tttcctattg acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg
     5041 tcaattacgg taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac
     5101 cattgacgtc aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga
     5161 cgtcaatgac ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg
     5221 caagtacatt actattggaa ggacgccagg gtacattggc agtactccca ttgacgtcaa
     5281 tggcggtaaa tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg
     5341 gcaatgacgc aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag
     5401 gtctatataa gcaatgctcg tttagggaac
//
