LOCUS       sequence-439159-        8900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of OPHN1 (pLxIS) in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8900)
  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 8900)
  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-OPHN1
FEATURES             Location/Qualifiers
     source          1..8900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      7..16
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             16..39
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2523..3089
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2539..3091
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2683..2700)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2910..2929
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3103..3105,3106..3108,3109..3822)
                     /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(3148..3169)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3409..3428)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3756..3777
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3884..4398
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4560..4576
                     /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(4584..4590,4591..4608,4609..4614))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4629..4650
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4767..4784)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4938..5526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5018..5037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5697..6488,6489..6557))
                     /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     6320..6339
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6558..6662)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6730..6748
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6786..6808)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6908..6927
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7121..7143
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7271..7290
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             7490..8006
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    8070..8411
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             8478..8894
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8824..8846
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8862..8878
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 agatctgcca ccatggacta caaagacgat gacgacaagg gcgtccaagt ggagacaatc
       61 tctcctgggg atggaaggac attccccaag cggggtcaga cctgcgttgt tcattatacc
      121 ggtatgttgg aagacggtaa aaaagtcgat agctcacggg atcgcaataa gcctttcaag
      181 ttcatgcttg ggaaacagga agtaatacgc ggctgggaag agggagtagc tcaaatgtcc
      241 gtcggtcaac gcgccaaatt gacaatttct cctgactacg cttatggtgc aaccggacac
      301 cccggcataa ttcccccaca cgcaacactt gtatttgatg tagaattgct caagctggaa
      361 acaagaggag tccaggttga aaccatatct cctggggacg gaagaacctt ccctaagcgg
      421 ggacaaactt gcgttgtcca ttacacaggc atgcttgaag atggtaagaa agtcgactca
      481 tcacgagaca gaaataagcc attcaaattt atgctcggta aacaggaagt cataagaggc
      541 tgggaagaag gagtggccca aatgtctgta ggacaacgag ccaaactcac aatatctccc
      601 gattatgctt atggtgccac tggccacccc ggcattatac cacctcatgc cactcttgtt
      661 tttgacgtag agctgttgaa gttggagacc cggggcgtgc aggttgagac tataagtcct
      721 ggagatggaa ggacttttcc taaaagaggt cagacatgcg tagtacacta tacagggatg
      781 ttggaggacg gcaaaaaagt cgacagctcc cgagaccgga ataagccttt taagttcatg
      841 ttggggaaac aggaagtaat ccgcgggtgg gaagagggag tcgctcagat gtctgtggga
      901 cagcgcgcta agctgaccat ttcccccgat tatgcctacg gcgctaccgg tcaccccggg
      961 attattccac ctcacgctac tttggtgttc gatgtcgaac ttctgaaact ggagactcgg
     1021 ggagtacagg ttgaaacaat cagcccagga gacggaagaa cctttccaaa aaggggtcaa
     1081 acatgtgtgg tacactacac aggaatgttg gaagacggaa aaaaggtgga cagctctcga
     1141 gaccgcaata aacctttcaa gtttatgctg ggcaaacagg aggtgatacg cggatgggag
     1201 gaaggcgtag cccagatgtc cgtgggtcag cgggcaaaac ttacaatttc acccgattac
     1261 gcatatggcg caacaggaca tccaggtatt ataccaccac atgccaccct cgttttcgac
     1321 gtagagcttt tgaaactcga aggggctgga gcaggcgcta tgggacaccc accactggaa
     1381 ttttcagatt gttatctcga ttctcccgat ttcagacaaa gactcaaata ctacgaagag
     1441 gaattggagc gcactaacaa gtttataaag gatgtaataa aggatgggtc cgccttgata
     1501 tctgctatga ggaactattc ttctgccgtt cagaaattct cacaaacact ccaaagcttc
     1561 cagttcgatt tcataggaga cacattgact gatgatgaaa taaatatcgc cgagtccttc
     1621 aaagaatttg ctgaactgtt gaatgaggtg gagaacgaga gaatgatgat ggtgcagaac
     1681 gccagcgacc tcctgattaa gcccctggaa accttccgca aagaacagat tggctttact
     1741 aaagagagga agaaaaagtt tgagaaggac ggcgagcgct tctattcatt gctcgacagg
     1801 catcttcatt tgtcaagtaa aaagaaagag agccaactct tggaggcaga tttgcaggta
     1861 gataaagaaa gacacaactt ttttgaaagt agtctggatt acgtatatca aatacaggat
     1921 gtccaggaaa gtaagaagtt taacattgtg gagccagtat tggctttcct ccatagcctt
     1981 ttcatatcca attctcttac tgtcgaattg actcaggact tcctccccta taaacaacag
     2041 ctccagctga gtttgcaaaa cacacgaaat cacttcagtt ctactcggga ggagatggag
     2101 gagctcaaaa aaaggatgaa agaggcccca caaacatgta aactcccagg tcagcctaca
     2161 atagaaggat atctctatac ccaagagaaa tgggctttgg gtattagctg ggccaagtat
     2221 tactgtaggt acgagaaaga gactcgaatg ttgacaatga ttcccatgga acaaaaaccc
     2281 ggtgcaaagc agggaccagt ggatctcaca cttaaatatt gtgtccgaag gaagactgaa
     2341 tccattgata agaggttttg tttcgacata gagaccaatg agcgccccgg taccataaca
     2401 ttgcaagctc cctccgaagc aaacagacgc ttgtggatgg aagccatgga tggtaaagag
     2461 cctatatatt gataagcggc cgcgtttaaa cgtcgacggt atcgataagc ttgatatcga
     2521 attccgcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     2581 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     2641 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     2701 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     2761 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     2821 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     2881 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     2941 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     3001 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     3061 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgag caagggcgag
     3121 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     3181 aagttcagcg tgtctggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     3241 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     3301 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     3361 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     3421 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     3481 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     3541 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggcgaacttc
     3601 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     3661 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     3721 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     3781 gccgccggga tcactctcgg catggacgag ctgtacaagt aatgaattaa ttaagaattc
     3841 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     3901 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg
     3961 agaatagaga agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca
     4021 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     4081 tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     4141 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     4201 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca
     4261 gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     4321 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     4381 tcagcggggg tctttcatgg gtaacagttt cttgaagttg gagaacaaca ttctgagggt
     4441 aggagtcgaa tattaagtaa tcctgactca attagccact gttttgaatc cacatactcc
     4501 aatactcctg aaatagttca ttatggacag cgcagaaaga gctggggaga attgtgaaat
     4561 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4621 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4681 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4741 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4801 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4861 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4921 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4981 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5041 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5101 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5161 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5221 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5281 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5341 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5401 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5461 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5521 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5581 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5641 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5701 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5761 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5821 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5881 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5941 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6001 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6061 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6121 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6181 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6241 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6301 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6361 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6421 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6481 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6541 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6601 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6661 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     6721 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     6781 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     6841 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     6901 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     6961 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     7021 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     7081 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     7141 acgacggcgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     7201 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     7261 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     7321 tgccggccac gatgcgtccg gcgtagaggc gattagtcca atttgttaaa gacaggatat
     7381 cagtggtcca ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg
     7441 agccatagat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     7501 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac
     7561 ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg
     7621 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     7681 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     7741 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     7801 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     7861 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt
     7921 gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc agattgattg
     7981 actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag acccctgccc
     8041 agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc gtgtctgtct
     8101 ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg tactagttag
     8161 ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga acacccggcc
     8221 gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg acctgaggaa
     8281 gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga gacgagaacc
     8341 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga agccgcgcgt
     8401 cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt
     8461 atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact
     8521 ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg
     8581 ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct
     8641 ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac
     8701 acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct
     8761 gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc
     8821 tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc
     8881 cttctctagg cgccggaatt
//
