LOCUS       sequence-462100-        8078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse STING amino acids 1-339 in mammalian cells. Stable 
            expression with piggybac transposase. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8078)
  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 8078)
  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 (1-339)
FEATURES             Location/Qualifiers
     source          1..8078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(101..120)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      914..1502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1403..1422
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1656..1673
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1790..1811
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1826..1831,1832..1849,1850..1856)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1864..1880
                     /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     1869..1891
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1888..1904
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1888..1904
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1925..1943
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2092..2126
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    2403..2423
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        2424..3602
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2655..3593
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3550..3570
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    3602..3626
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4653..4677)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4755..4774)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4820..4875
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4874..4893)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        5230..5609
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5610..5813
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5764..5784
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5846..6445
                     /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(5846..5865)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6342..6362
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6483..6500)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6489..6713
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7063..7125
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7191..7210)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7192..7210)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7217..7234)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7217..7233)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7226..7248)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7375..7830
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7462..7481)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7672..7693
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7856..7960
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7961..8029,8030..8078,1..743)
                     /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"
ORIGIN
        1 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
       61 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      121 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      181 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
      241 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
      301 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
      361 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
      421 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
      481 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
      541 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
      601 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
      661 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
      721 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
      781 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
      841 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
      901 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgccgcttg caaacaaaaa
      961 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1021 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1081 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1141 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1201 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1261 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1321 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1381 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1441 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1501 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1561 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     1621 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     1681 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     1741 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     1801 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     1861 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     1921 tcgaaattaa ccctcactaa agggaacaaa agctggtacc tcgcgcgact tggtttgcca
     1981 ttctttagcg cgcgtcgcgt cacacagctt ggccacaatg tggtttttgt caaacgaaga
     2041 ttctatgacg tgtttaaagt ttaggtcgag taaagcgcaa atctttttta accctagaaa
     2101 gatagtctgc gtaaaattga cgcatgcatt cttgaaatat tgctctctct ttctaaatag
     2161 cgcgaatccg tcgctgtgca tttaggacat ctcagtcgcc gcttggagct cccgtgaggc
     2221 gtgcttgtca atgcggtaag tgtcactgat tttgaactat aacgaccgcg tgagtcaaaa
     2281 tgacgcatga ttatctttta cgtgactttt aagatttaac tcatacgata attatattgt
     2341 tatttcatgt tctacttacg tgataactta ttatatatat attttcttgt tatagatatc
     2401 atcaactttg tatagaaaag ttgggctccg gtgcccgtca gtgggcagag cgcacatcgc
     2461 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
     2521 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     2581 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     2641 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
     2701 atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg
     2761 agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc
     2821 ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc
     2881 accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac
     2941 ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca
     3001 ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat
     3061 gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag
     3121 ctggccggcc tgctctggtg cctggtctcg cgccgccgtg tatcgccccg ccctgggcgg
     3181 caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg
     3241 ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca
     3301 cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc
     3361 gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt
     3421 ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta
     3481 ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt
     3541 ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt
     3601 gacaagtttg tacaaaaaag caggctgcca ccatgccata ctccaacctg catccagcca
     3661 tcccacggcc cagaggtcac cgctccaaat atgtagccct catctttctg gtggccagcc
     3721 tgatgatcct ttgggtggca aaggatccac caaatcacac tctgaagtac ctagcacttc
     3781 acctagcctc gcacgaactt ggactactgt tgaaaaacct ctgctgtctg gctgaagagc
     3841 tgtgccatgt ccagtccagg taccagggca gctactggaa ggctgtgcgc gcctgcctgg
     3901 gatgccccat ccactgtatg gctatgattc tactatcgtc ttatttctat ttcctccaaa
     3961 acactgctga catatacctc agttggatgt ttggccttct ggtcctctat aagtccctaa
     4021 gcatgctcct gggccttcag agcttgactc cagcggaagt ctctgcagtc tgtgaagaaa
     4081 agaagttaaa tgttgcccac gggctggcct ggtcatacta cattgggtac ttgcggttga
     4141 tcttaccagg gctccaggcc cggatccgaa tgttcaatca gctacataac aacatgctca
     4201 gtggtgcagg gagccgaaga ctgtacatcc tctttccatt ggactgtggg gtgcctgaca
     4261 acctgagtgt agttgacccc aacattcgat tccgagatat gctgccccag caaaacatcg
     4321 accgtgctgg catcaagaat cgggtttatt ccaacagcgt ctacgagatt ctggagaacg
     4381 gacagccagc aggcgtctgt atcctggagt acgccacccc cttgcagacc ctgtttgcca
     4441 tgtcacagga tgccaaagct ggcttcagtc gggaggatcg gcttgagcag gctaaactct
     4501 tctgccggac acttgaggaa atcctggaag atgtccccga gtctcgaaat aactgccgcc
     4561 tcattgtcta ccaagaaccc acagacggaa acagtttctc actgtctcag gaggtgctcc
     4621 ggcacattcg tcaggaagaa aaggaggagt gaacccagct ttcttgtaca aagtggtgat
     4681 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     4741 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     4801 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     4861 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     4921 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     4981 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     5041 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     5101 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     5161 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     5221 cttcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     5281 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     5341 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     5401 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     5461 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     5521 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     5581 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     5641 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     5701 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     5761 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
     5821 taactagaga acccactgcg ccaccatgac cgagtacaag cccacggtgc gcctcgccac
     5881 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     5941 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     6001 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     6061 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     6121 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     6181 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     6241 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     6301 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     6361 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     6421 catgacccgc aagcccggtg cctgactcga gtctagaggg cccgtttaaa cccgctgatc
     6481 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     6541 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     6601 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     6661 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggctcgagt
     6721 taattaacga gagcataata ttgatatgtg ccaaagttgt ttctgactga ctaataagta
     6781 taatttgttt ctattatgta taggttaagc taattactta ttttataata caacatgact
     6841 gtttttaaag tacaaaataa gtttattttt gtaaaagaga gaatgtttaa aagttttgtt
     6901 actttataga agaaattttg agtttttgtt tttttttaat aaataaataa acataaataa
     6961 attgtttgtt gaatttatta ttagtatgta agtgtaaata taataaaact taatatctat
     7021 tcaaattaat aaataaacct cgatatacag accgataaaa cacatgcgtc aattttacgc
     7081 atgattatct ttaacgtacg tcacaatatg attatctttc tagggttaaa taatagtttc
     7141 taattttttt attattcagc ctgctgtcgt gaataccgag ctccaattcg ccctatagtg
     7201 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     7261 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     7321 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     7381 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     7441 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     7501 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     7561 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     7621 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     7681 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     7741 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     7801 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     7861 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     7921 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     7981 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     8041 gctggtgaaa gtaaaagatg ctgaagatca gttgggtg
//
