LOCUS       sequence-461196-        7332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of cyclic GMP/AMP synthase (cGAS), as a nuclear rupture 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7332)
  AUTHORS   Korsten G, Osinga M, Pelle RA, Serweta AK, Hoogenberg B, Kampinga 
            HH, Kapitein LC
  TITLE     Nuclear poly-glutamine aggregates rupture the nuclear envelope and 
            hinder its repair.
  JOURNAL   J Cell Biol. 2024 Nov 4;223(11):e202307142. doi: 
            10.1083/jcb.202307142. Epub 2024 Aug 16.
  PUBMED    39150509
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: "J Cell 
            Biol."; date: "2024-11-4"; volume: "223(11):e202307142. doi"; pages:
            " 10.1083/jcb.202307142. Epub 2024 Aug 16"
COMMENT     Sequence Label: mCherry-cGAS-HA
FEATURES             Location/Qualifiers
     source          1..7332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1981..2000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2172..2551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2552..2755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2706..2726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2757..2775
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2778..2796
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2806..2830
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2921..3628
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3066..3084)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3334..3353
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5210..5236
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5277..5294)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5283..5507
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5553..5981
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5640..5659)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5850..5871
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5990..6010)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5995..6324
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6175..6310
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6372..6419
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6438..6812
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    6942..7063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6979..6998)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7033..7052
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7112..7128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7112..7128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7125..7147)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7136..7152
                     /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(7160..7166,7167..7184,7185..7190))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7205..7226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctccc
     2761 tatcagtgat agagatctcc ctatcagtga tagagatcgt cgacgagctc gtttagtgaa
     2821 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
     2881 ccgatccagc ctccggactc tagcgtttaa actaaagctt atggtgagca agggcgagga
     2941 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     3001 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     3061 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     3121 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     3181 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     3241 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     3301 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     3361 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     3421 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     3481 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     3541 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     3601 caccggcggc atggacgagc tgtacaagct cgagatgcag ccttggcacg gaaaggccat
     3661 gcagagagct tccgaggccg gagccactgc ccccaaggct tccgcacgga atgccagggg
     3721 cgccccgatg gatcccaccg agtctccggc tgcccccgag gccgccctgc ctaaggcggg
     3781 aaagttcggc cccgccagga agtcgggatc ccggcagaaa aagagcgccc cggacaccca
     3841 ggagaggccg cccgtccgcg caactggggc ccgcgccaaa aaggcccctc agcgcgccca
     3901 ggacacgcag ccgtctgacg ccaccagcgc ccctggggca gaggggctgg agcctcctgc
     3961 ggctcgggag ccggctcttt ccagggctgg ttcttgccgc cagaggggcg cgcgctgctc
     4021 cacgaagcca agacctccgc ccgggccctg ggacgtgccc agccccggcc tgccggtctc
     4081 ggcccccatt ctcgtacgga gggatgcggc gcctggggcc tcgaagctcc gggcggtttt
     4141 ggagaagttg aagctcagcc gcgatgatat ctccacggcg gcggggatgg tgaaaggggt
     4201 tgtggaccac ctgctgctca gactgaagtg cgactccgcg ttcagaggcg tcgggctgct
     4261 gaacaccggg agctactatg agcacgtgaa gatttctgca cctaatgaat ttgatgtcat
     4321 gtttaaactg gaagtcccca gaattcaact agaagaatat tccaacactc gtgcatatta
     4381 ctttgtgaaa tttaaaagaa atccgaaaga aaatcctctg agtcagtttt tagaaggtga
     4441 aatattatca gcttctaaga tgctgtcaaa gtttaggaaa atcattaagg aagaaattaa
     4501 cgacattaaa gatacagatg tcatcatgaa gaggaaaaga ggagggagcc ctgctgtaac
     4561 acttcttatt agtgaaaaaa tatctgtgga tataaccctg gctttggaat caaaaagtag
     4621 ctggcctgct agcacccaag aaggcctgcg cattcaaaac tggctttcag caaaagttag
     4681 gaagcaacta cgactaaagc cattttacct tgtacccaag catgcaaagg aaggaaatgg
     4741 tttccaagaa gaaacatggc ggctatcctt ctctcacatc gaaaaggaaa ttttgaacaa
     4801 tcatggaaaa tctaaaacgt gctgtgaaaa caaagaagag aaatgttgca ggaaagattg
     4861 tttaaaacta atgaaatacc ttttagaaca gctgaaagaa aggtttaaag acaaaaaaca
     4921 tctggataaa ttctcttctt atcatgtgaa aactgccttc tttcacgtat gtacccagaa
     4981 ccctcaagac agtcagtggg accgcaaaga cctgggcctc tgctttgata actgcgtgac
     5041 atactttctt cagtgcctca ggacagaaaa acttgagaat tattttattc ctgaattcaa
     5101 tctattctct agcaacttaa ttgacaaaag aagtaaagaa tttctgacaa agcaaattga
     5161 atatgaaaga aacaatgagt ttccagtttt tgatgaattt agcggccgct acccatacga
     5221 tgttccagat tacgcttgac tcgagtctag agggcccgtt taaacccgct gatcagcctc
     5281 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     5341 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     5401 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     5461 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     5521 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     5581 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     5641 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5701 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5761 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5821 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5881 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5941 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     6001 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6061 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     6121 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     6181 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     6241 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     6301 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     6361 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     6421 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
     6481 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
     6541 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
     6601 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
     6661 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
     6721 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
     6781 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
     6841 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6901 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6961 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     7021 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     7081 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     7141 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     7201 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     7261 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     7321 gtattgggcg ct
//
