LOCUS       sequence-462106-        8195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse STING in mammalian cells. Stable expression with 
            piggybac transposase. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8195)
  AUTHORS   Bentley-DeSousa A, Roczniak-Ferguson A, Ferguson SM
  TITLE     A STING-CASM-GABARAP pathway activates LRRK2 at lysosomes.
  JOURNAL   J Cell Biol. 2025 Feb 3;224(2):e202310150. doi: 
            10.1083/jcb.202310150. Epub 2025 Jan 15.
  PUBMED    39812709
REFERENCE   2  (bases 1 to 8195)
  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: "2025-02-3"; volume: "224(2):e202310150. doi"; pages: 
            " 10.1083/jcb.202310150. Epub 2025 Jan 15"
COMMENT     Sequence Label: pPB-EF1A-mSTING
FEATURES             Location/Qualifiers
     source          1..8195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..39
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    316..336
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        337..1515
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          568..1506
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1463..1483
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1515..1539
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2683..2707)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2785..2804)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2850..2905
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2904..2923)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        3260..3639
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3640..3843
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3794..3814
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3876..4475
                     /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(3876..3895)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4372..4392
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4513..4530)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4519..4743
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5093..5155
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5221..5240)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5222..5240)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5247..5264)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5247..5263)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5256..5278)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5405..5860
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5492..5511)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5702..5723
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5886..5990
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5991..6059,6060..6851)
                     /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     complement(6209..6228)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7022..7610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7511..7530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7764..7781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7898..7919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7934..7939,7940..7957,7958..7964)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7972..7988
                     /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)."
     primer_bind     7977..7999
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7996..8012
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7996..8012
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8033..8051
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc
       61 tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga
      121 gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc
      181 gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg
      241 ataattatat tgttatttca tgttctactt acgtgataac ttattatata tatattttct
      301 tgttatagat atcatcaact ttgtatagaa aagttgggct ccggtgcccg tcagtgggca
      361 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt
      421 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
      481 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
      541 cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc
      601 ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg
      661 attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa
      721 ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg
      781 cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa
      841 aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc
      901 caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg
      961 tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg
     1021 gggtagtctc aagctggccg gcctgctctg gtgcctggtc tcgcgccgcc gtgtatcgcc
     1081 ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg
     1141 cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg
     1201 ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac
     1261 tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg
     1321 tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg
     1381 gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt
     1441 gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca
     1501 tttcaggtgt cgtgacaagt ttgtacaaaa aagcaggctg ccaccatgcc atactccaac
     1561 ctgcatccag ccatcccacg gcccagaggt caccgctcca aatatgtagc cctcatcttt
     1621 ctggtggcca gcctgatgat cctttgggtg gcaaaggatc caccaaatca cactctgaag
     1681 tacctagcac ttcacctagc ctcgcacgaa cttggactac tgttgaaaaa cctctgctgt
     1741 ctggctgaag agctgtgcca tgtccagtcc aggtaccagg gcagctactg gaaggctgtg
     1801 cgcgcctgcc tgggatgccc catccactgt atggctatga ttctactatc gtcttatttc
     1861 tatttcctcc aaaacactgc tgacatatac ctcagttgga tgtttggcct tctggtcctc
     1921 tataagtccc taagcatgct cctgggcctt cagagcttga ctccagcgga agtctctgca
     1981 gtctgtgaag aaaagaagtt aaatgttgcc cacgggctgg cctggtcata ctacattggg
     2041 tacttgcggt tgatcttacc agggctccag gcccggatcc gaatgttcaa tcagctacat
     2101 aacaacatgc tcagtggtgc agggagccga agactgtaca tcctctttcc attggactgt
     2161 ggggtgcctg acaacctgag tgtagttgac cccaacattc gattccgaga tatgctgccc
     2221 cagcaaaaca tcgaccgtgc tggcatcaag aatcgggttt attccaacag cgtctacgag
     2281 attctggaga acggacagcc agcaggcgtc tgtatcctgg agtacgccac ccccttgcag
     2341 accctgtttg ccatgtcaca ggatgccaaa gctggcttca gtcgggagga tcggcttgag
     2401 caggctaaac tcttctgccg gacacttgag gaaatcctgg aagatgtccc cgagtctcga
     2461 aataactgcc gcctcattgt ctaccaagaa cccacagacg gaaacagttt ctcactgtct
     2521 caggaggtgc tccggcacat tcgtcaggaa gaaaaggagg aggttaccat gaatgccccc
     2581 atgacctcag tggcacctcc tccctccgta ctgtcccaag agccaagact cctcatcagt
     2641 ggtatggatc agcctctccc actccgcact gacctcatct gaacccagct ttcttgtaca
     2701 aagtggtgat cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     2761 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     2821 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     2881 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     2941 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
     3001 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     3061 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     3121 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     3181 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct
     3241 gcagcccaag cttcgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     3301 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     3361 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     3421 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     3481 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     3541 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     3601 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     3661 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     3721 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     3781 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     3841 gctctctggc taactagaga acccactgcg ccaccatgac cgagtacaag cccacggtgc
     3901 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     3961 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     4021 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4081 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4141 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4201 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4261 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4321 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     4381 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     4441 gcacctggtg catgacccgc aagcccggtg cctgactcga gtctagaggg cccgtttaaa
     4501 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     4561 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     4621 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     4681 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     4741 tggctcgagt taattaacga gagcataata ttgatatgtg ccaaagttgt ttctgactga
     4801 ctaataagta taatttgttt ctattatgta taggttaagc taattactta ttttataata
     4861 caacatgact gtttttaaag tacaaaataa gtttattttt gtaaaagaga gaatgtttaa
     4921 aagttttgtt actttataga agaaattttg agtttttgtt tttttttaat aaataaataa
     4981 acataaataa attgtttgtt gaatttatta ttagtatgta agtgtaaata taataaaact
     5041 taatatctat tcaaattaat aaataaacct cgatatacag accgataaaa cacatgcgtc
     5101 aattttacgc atgattatct ttaacgtacg tcacaatatg attatctttc tagggttaaa
     5161 taatagtttc taattttttt attattcagc ctgctgtcgt gaataccgag ctccaattcg
     5221 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     5281 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     5341 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     5401 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5461 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5521 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5581 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5641 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5701 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5761 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5821 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     5881 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5941 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6001 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6061 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6121 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6181 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6241 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6301 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6361 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6421 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6481 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6541 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6601 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6661 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6721 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6781 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6841 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6901 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6961 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7021 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7081 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7141 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7201 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7261 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7321 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7381 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7441 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7501 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7561 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7621 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7681 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     7741 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     7801 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     7861 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     7921 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     7981 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc gaaattaacc
     8041 ctcactaaag ggaacaaaag ctggtacctc gcgcgacttg gtttgccatt ctttagcgcg
     8101 cgtcgcgtca cacagcttgg ccacaatgtg gtttttgtca aacgaagatt ctatgacgtg
     8161 tttaaagttt aggtcgagta aagcgcaaat ctttt
//
