LOCUS       sequence-440567-        7886 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of TASL pLxIS-STING PLPLR in mammalian cells by 
            retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7886)
  AUTHORS   Landau LM, Chaudhary N, Tien YC, Rogozinska M, Joshi S, Yao C, 
            Crowley J, Hullahalli K, Campbell IW, Waldor MK, Haigis M, Kagan JC
  TITLE     pLxIS-containing domains are biochemically flexible regulators of 
            interferons and metabolism.
  JOURNAL   Mol Cell. 2024 Jul 11;84(13):2436-2454.e10. doi: 
            10.1016/j.molcel.2024.05.030. Epub 2024 Jun 25.
  PUBMED    38925114
REFERENCE   2  (bases 1 to 7886)
  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: 
            "10.1016/j.molcel.2024.05.030"; journalName: "Mol Cell"; date: 
            "2024-07-11- 11"; volume: "84"; issue: "13"; pages: "2436-2454.e10"
COMMENT     Sequence Label: pMSCV-FKBP-TASL(pLxIS-PLPLR)
FEATURES             Location/Qualifiers
     source          1..7886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(494..513)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(641..663)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(857..876)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     976..998
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1036..1054)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1122..1226
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1227..1295,1296..2087)
                     /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(1445..1464)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2258..2846
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2747..2766
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3000..3017
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3134..3155
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3170..3175,3176..3193,3194..3200)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3208..3224
                     /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)."
     LTR             3386..3900
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(3962..4675,4676..4678,4679..4681))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4007..4028)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4356..4375
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4615..4636
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    4693..5245
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4695..5261
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4855..4874)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     5084..5101
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(6731..6754)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      6754..6763
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(6776..7192)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(6792..6808)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(6824..6846)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    7259..7600
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             join(7664..7886,1..294)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tggttagttc aaataaggca cagggtcatt tcaggtcctt ggggcaccct ggaaacatct
       61 gatggttctc tagaaactgc tgagggcggg accgcatctg gggaccatct gttcttggcc
      121 ctgagccggg gcaggaactg cttaccacag atatcctgtt tggcccatat tctgctgtct
      181 ctctgttcct aaccttgatc tgaacttctc tattctcagt tatgtatttt ccatgccttg
      241 caaaatggcg ttacttaagc tagcttgcca aacctacagg tggggtcttt cattcccccc
      301 tttttctgga gactaaataa aatcttttat tttatctatg gctcgtactc tataggcttc
      361 agctggtgat attgttgagt caaaactaga gcctggacca ctgatatcct gtctttaaca
      421 aattggacta atcgcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
      481 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
      541 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
      601 ttgggcgcca tctccttgca tgcaccattc cttgcgccgt cgttttacaa cgtcgtgact
      661 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
      721 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
      781 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
      841 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
      901 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
      961 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     1021 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     1081 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1141 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1201 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1261 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1321 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     1381 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1441 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     1501 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1561 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1621 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1681 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1741 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1801 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1861 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1921 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1981 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2041 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2101 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2161 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2221 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2281 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2341 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2401 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2461 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2521 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2581 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2641 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2701 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2761 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2821 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2881 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2941 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3001 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     3061 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     3121 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     3181 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac aattctcccc
     3241 agctctttct gcgctgtcca taatgaacta tttcaggagt attggagtat gtggattcaa
     3301 aacagtggct aattgagtca ggattactta atattcgact cctaccctca gaatgttgtt
     3361 ctccaacttc aagaaactgt tacccatgaa agacccccgc tgacgggtag tcaatcactc
     3421 agaggagacc ctcccaagga acagcgagac cacaagtcgg atgcaactgc aagagggttt
     3481 attggataca cgggtacccg ggcgacgcag tctatcggag gactggcgcg ccgagtgagg
     3541 ggttgtgggc tcttttattg agctcgggga gcagaagcgc gcgaacagaa gcgagaagcg
     3601 aactgattgg ttagttcaaa taaggcacag ggtcatttca ggtccttggg gcaccctgga
     3661 aacatctgat ggttctctag aaactgctga gggcgggacc gcatctgggg accatctgtt
     3721 cttggccctg agccggggca ggaactgctt accacagata tcctgtttgg cccatattct
     3781 gctgtctctc tgttcctaac cttgatctga acttctctat tctcagttat gtattttcca
     3841 tgccttgcaa aatggcgtta cttaagctag cttgccaaac ctacaggtgg ggtctttcat
     3901 tccccccttt ttctggagac taaataaaat cttttatttt atcgaattct taattaattc
     3961 attacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca
     4021 ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag ggcggactgg gtgctcaggt
     4081 agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt
     4141 cggcgagctg cacgctgccg tcctcgatgt tgtggcggat cttgaagttc gccttgatgc
     4201 cgttcttctg cttgtcggcc atgatataga cgttgtggct gttgtagttg tactccagct
     4261 tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca
     4321 ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga
     4381 agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct
     4441 gcttcatgtg gtcggggtag cggctgaagc actgcacgcc gtaggtcagg gtggtcacga
     4501 gggtgggcca gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc
     4561 cgtaggtggc atcgccctcg ccctcgccag acacgctgaa cttgtggccg tttacgtcgc
     4621 cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca
     4681 tggttgtggc catattatca tcgtgttttt caaaggaaaa ccacgtcccc gtggttcggg
     4741 gggcctagac gtttttttaa cctcgactaa acacatgtaa agcatgtgca ccgaggcccc
     4801 agatcagatc ccatacaatg gggtaccttc tgggcatcct tcagcccctt gttgaatacg
     4861 cttgaggaga gccatttgac tctttccaca actatccaac tcacaacgtg gcactggggt
     4921 tgtgccgcct ttgcaggtgt atcttataca cgtggctttt ggccgcagag gcacctgtcg
     4981 ccaggtgggg ggttccgctg cctgcaaagg gtcgctacag acgttgtttg tcttcaagaa
     5041 gcttccagag gaactgcttc cttcacgaca ttcaacagac cttgcattcc tttggcgaga
     5101 ggggaaagac ccctaggaat gctcgtcaag aagacagggc caggtttccg ggccctcaca
     5161 ttgccaaaag acggcaatat ggtggaaaat aacatataga caaacgcaca ccggccttat
     5221 tccaagcggc ttcggccagt aacgttaggg gggggggcgg aattcgatat caagcttatc
     5281 gataccgtcg acgtttaaac gcggccgctt atcaaatcag gtcggtcctg aggggcaagg
     5341 gttgatccat accgggtgtg atattactat attggcttat atgcagggat ggggtgctaa
     5401 tttcagatga agccccagcc ccagcaccct caagcttcag aagctcgaca tcgaatacca
     5461 gggtagcgtg gggtggaatt atgccagggt ggcccgtcgc gccgtaggca taatcaggag
     5521 agatagtcag ttttgctctc tggcccacgc tcatctgtgc cacaccttcc tcccagccgc
     5581 gaatcacctc ctgctttccc aacatgaatt taaacggttt gttgcggtcg cgtgacgaat
     5641 cgaccttttt cccatcttct aacatccccg tatagtgcac gacacacgtc tggcctcttt
     5701 tggggaatgt tctcccatcc cctggactga ttgtttccac ttgtacaccg cgcgtctcta
     5761 actttaaaag ttccacatcg aacactaagg ttgcgtgagg tggtatgatc cctgggtgac
     5821 cggtggctcc gtaggcgtag tctggactga tggttaattt ggcccgttgt cccactgaca
     5881 tctgagctac gccttcttcc caacctctga tgacttcctg cttcccaagc ataaacttaa
     5941 agggtttgtt gcgatcacga cttgagtcaa ctttcttacc atcctccaac atcccagtgt
     6001 agtggacaac acaggtctgt ccccgctttg ggaaagttct tccgtcaccg ggcgaaatag
     6061 tctctacctg caccccccgc gtctcaagct taagtaactc gacgtcgaag acaagtgtcg
     6121 cgtgcggcgg aatgatacca gggtgcccag tcgcaccgta cgcgtaatca ggacttattg
     6181 tcagtttagc tctttgcccg acagacattt gcgccacacc ctcttcccat ccgcgtatta
     6241 cttcctgttt tccaagcatg aatttgaagg gtttatttct atcgcggctt gaatcgactt
     6301 tcttgccgtc ttccagcata ccggtgtaat gcacgacgca cgtttgtccc cgtttaggaa
     6361 aggtgcgtcc atcacctggt gaaatcgttt cgacttgaac tccccgcgtc tccaacttca
     6421 acagctctac atcaaacaca agtgtcgcat gcgggggaat gatcccagga tggcctgtag
     6481 ccccatatgc gtaatctggc gagatggtta acttagcacg ctgcccaacg gacatttggg
     6541 ccacgccttc ctcccaacca cggataacct cctgtttccc cagcataaat ttgaaaggtt
     6601 tgttacggtc tctagagcta tcgaccttct taccgtcctc cagcatgccg gtataatgga
     6661 ctacgcaagt ttggccacgc ttaggaaatg tacgcccgtc tcctggagat attgtttcta
     6721 cttgaacgcc tttgtcgtca tcgtccttgt aatccatggt ggcagatcta attccggcgc
     6781 ctagagaagg agtgagggct ggataaaggg aggatcgagg cggggtcgaa cgaggaggtt
     6841 caagggggag agacggggcg gatggaggaa gaggaggcgg aggcttaggg tgtacaaagg
     6901 gcttgaccca gggagggggg tcaaaagcca aggcttccca ggtcacgatg taggggacct
     6961 ggtctgggtg tccatgcggg ccaggtgaaa agaccttgat cttaacctgg gtgatgaggt
     7021 ctcggttaaa ggtgccgtct cgcggccatc cgacgttaaa ggttggccat tctgcagagc
     7081 agaaggtaac ccaacgtctc ttcttgacat ctaccgactg gttgtgagcg atccgctcga
     7141 catctttcca gtgacctaag gtcaaactta agggagtggt aacagtctgg ccctaatttt
     7201 cagacaaata cagaaacaca gtcagacaga gacaacacag aacgatgctg cagcgctgca
     7261 gcagacaaga cgcgcggctt cggttccaaa ccgaaagcaa aaattcagac ggaggcggga
     7321 actgttttag gttctcgtct cctaccagaa ccacatatcc tgacggggtc ggattccaca
     7381 tcgactccct tcctcaggtc gggccacaaa aacggccccc aaagtccctg ggacgtctcc
     7441 cagggttgcg gccgggtgtt cagaactcgt cagttccacc acgggtccgc cagatacaga
     7501 gctagttagc taactagtac agacgcaggc gcaaacatta gatgccggca caaacacgca
     7561 caaagacaga gacagacacg aaacgaccgc tggccagctt acctcccggc gggggggtcg
     7621 gtggtccctg ggcaggggtc tccaaatctc ggtggaacct ccaaatgaaa gacccccgag
     7681 gtgggcagtc aatcaatctg aggagaccct cccaaggatc agcgagtcca cgattcggat
     7741 gcaaacagca agaggcttta ttgggaatac gggtacccgg gcgacgcagt ctatcggagg
     7801 actggcgcgc cgagtgaggg gttgtgggct cttttattga gctcggggag cagaagcgcg
     7861 cgaacagaag cgagaagcga actgat
//
