LOCUS       sequence-438524-        8465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of RFP (pLxIS-PLPLR) in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8465)
  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 8465)
  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-RFP(pLxIS-PLPLR)
FEATURES             Location/Qualifiers
     source          1..8465
                     /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    2088..2654
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2104..2656
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2248..2265)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2475..2494
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2668..2670,2671..2673,2674..3387)
                     /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(2713..2734)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2974..2993)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3321..3342
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3449..3963
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4125..4141
                     /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(4149..4155,4156..4173,4174..4179))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4194..4215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4332..4349)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4503..5091)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4583..4602)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5262..6053,6054..6122))
                     /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     5885..5904
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6123..6227)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6295..6313
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6351..6373)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6473..6492
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6686..6708
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6836..6855
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             7055..7571
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7635..7976
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             8043..8459
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8389..8411
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8427..8443
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 agatctgcca ccatggatta caaagacgac gatgataaag gagtgcaggt tgagacaata
       61 tcacctgggg acggccgcac attcccaaag agggggcaaa catgtgtggt gcactatact
      121 ggaatgctcg aagacggcaa aaaagttgat agttccagag atcgcaacaa gccctttaag
      181 ttcatgctgg gtaaacagga ggtcataaga ggctgggagg agggagttgc acaaatgagc
      241 gtgggtcaaa gggctaaact cactatttct ccagattacg catatggagc cacaggccac
      301 cctggcatca ttccccctca cgcaaccctg gtctttgacg ttgagctcct caagctcgaa
      361 actagaggtg tgcaggtgga gaccataagc cccggagatg gtagaacttt tcctaaaagg
      421 gggcaaactt gtgtggtgca ttataccgga atgctggaag acggaaaaaa ggtagactcc
      481 tcaagagata gaaacaagcc attcaaattc atgctcggaa agcaggaggt catcagaggc
      541 tgggaagagg gggtggctca gatgtctgtg ggacaacgcg ctaagttgac tattagccct
      601 gactacgcct atggtgctac tgggcaccct ggaatcatcc ccccacacgc tacccttgta
      661 tttgacgtag agttgttgaa acttgaaacc aggggtgtgc aagtagaaac aataagtcca
      721 ggtgatggca ggactttccc aaagcgcggc cagacatgtg tcgtccatta cactggtatg
      781 ttggaagatg gtaagaaggt agactctagc cgagatcgca acaaaccttt caaatttatg
      841 cttgggaagc aagaggttat ccgaggctgg gaggaaggtg tcgcccagat gtccgtcgga
      901 cagagggcaa aactgacaat aagtcccgac tacgcatacg gggctaccgg gcacccagga
      961 atcatccctc cccatgctac tctggtattt gacgtggagc tgcttaagct tgaaacacgc
     1021 ggtgttcagg tagaaacaat ctccccaggc gatggacgga ctttccctaa gagagggcaa
     1081 acatgtgtag tacattacac aggcatgctg gaggatggta aaaaagttga ttctagcagg
     1141 gaccggaaca agcctttcaa gttcatgttg ggaaaacagg aagtcatccg agggtgggag
     1201 gagggcgtag cacagatgtc cgttggacag cgcgcaaagc tgacaatctc ccctgattat
     1261 gcatacggcg caactggaca tcctgggatc atacctcccc atgccacctt ggtctttgac
     1321 gtagaactgt tgaagttgga aggtgctgga gcaggggcaa tgagaagcag caagaacgtt
     1381 atcaaggaat tcatgcggtt caaccttgtg atttcaggaa cagtaaatgg ccacccactg
     1441 ccattgcggg gagagggcga aggacgacct tacgaaggcc acaacacagt gaaacttaaa
     1501 gtcacaaagg gaggtccact gcctttcgcc tgggatatcc tttcacccca attccaatat
     1561 ggtagtaagg tttatgtaaa acaccccgct gatatccccg actataagaa actgtctttt
     1621 ccagagggat ttaagtggga gagagtgatg aattttgagg acgggggtgt cgtcacagtg
     1681 acccaagaca gttcccttca ggacggctgt tttatttata aggtaaagtt cattggggtc
     1741 aactttccta gcgatggccc agtcatgcag aaaaagacta tgggatggga ggcttctacc
     1801 gagaggcttt acccaagaga tggcgtcctc aagggggaaa tccataaggc tctcaaactg
     1861 aaggatgggg gccattacct tgtcgaattt aagagtatct acatggccaa aaaacccgta
     1921 cagttgccag gctattacta tgttgatagc aaattggata ttacttctca taatgaagac
     1981 tacacaattg ttgaacaata tgagaggaca gaagggagac atcatttgtt cctctgataa
     2041 gcggccgcgt ttaaacgtcg acggtatcga taagcttgat atcgaattcc gccccccccc
     2101 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     2161 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     2221 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     2281 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     2341 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     2401 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     2461 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     2521 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     2581 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     2641 aaacacgatg ataatatggc cacaaccatg gtgagcaagg gcgaggagct gttcaccggg
     2701 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtct
     2761 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     2821 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     2881 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     2941 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     3001 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3061 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     3121 tatatcatgg ccgacaagca gaagaacggc atcaaggcga acttcaagat ccgccacaac
     3181 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3241 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     3301 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3361 ctcggcatgg acgagctgta caagtaatga attaattaag aattcgataa aataaaagat
     3421 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     3481 agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc
     3541 agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa
     3601 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc
     3661 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     3721 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     3781 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact
     3841 gcgtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     3901 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     3961 catgggtaac agtttcttga agttggagaa caacattctg agggtaggag tcgaatatta
     4021 agtaatcctg actcaattag ccactgtttt gaatccacat actccaatac tcctgaaata
     4081 gttcattatg gacagcgcag aaagagctgg ggagaattgt gaaattgtta tccgctcaca
     4141 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4201 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4261 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4321 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4381 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4441 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4501 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4561 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4621 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4681 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4741 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4801 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4861 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4921 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4981 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5041 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5101 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5161 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5221 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5281 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5341 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5401 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5461 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5521 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5581 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5641 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5701 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5761 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5821 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5881 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5941 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6001 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6061 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6121 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6181 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6241 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     6301 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     6361 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     6421 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     6481 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     6541 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     6601 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     6661 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga
     6721 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     6781 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     6841 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     6901 gtccggcgta gaggcgatta gtccaatttg ttaaagacag gatatcagtg gtccaggctc
     6961 tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca tagataaaat
     7021 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     7081 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     7141 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
     7201 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     7261 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     7321 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     7381 tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
     7441 atagactgcg tcgcccgggt acccgtattc ccaataaagc ctcttgctgt ttgcatccga
     7501 atcgtggact cgctgatcct tgggagggtc tcctcagatt gattgactgc ccacctcggg
     7561 ggtctttcat ttggaggttc caccgagatt tggagacccc tgcccaggga ccaccgaccc
     7621 ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc tttgtgcgtg
     7681 tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta gttagctaac tagctctgta
     7741 tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac
     7801 gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga
     7861 atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc
     7921 ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc
     7981 gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt
     8041 agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc
     8101 ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg
     8161 cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca
     8221 tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc
     8281 cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg
     8341 tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc
     8401 ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccg
     8461 gaatt
//
