LOCUS       sequence-439087-        8090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of p27 (pLxIS-PLPLR) in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8090)
  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 8090)
  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-p27(pLxIS-PLPLR)
FEATURES             Location/Qualifiers
     source          1..8090
                     /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    1713..2279
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1729..2281
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1873..1890)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2100..2119
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2293..2295,2296..2298,2299..3012)
                     /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(2338..2359)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2599..2618)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2946..2967
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3074..3588
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3750..3766
                     /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(3774..3780,3781..3798,3799..3804))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3819..3840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3957..3974)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4128..4716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4208..4227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4887..5678,5679..5747))
                     /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     5510..5529
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5748..5852)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5920..5938
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5976..5998)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6098..6117
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6311..6333
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6461..6480
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6680..7196
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7260..7601
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7668..8084
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8014..8036
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8052..8068
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 agatctgcca ccatggatta taaggatgac gacgataagg gggtccaggt ggaaaccata
       61 tcacccggcg acggccgcac ttttcctaag cgggggcaga cctgcgtagt gcactacacc
      121 ggaatgcttg aagatggtaa aaaagttgat agttcccgcg accgaaataa gcctttcaaa
      181 tttatgctcg ggaaacaaga agtcatcaga ggttgggaag aaggagttgc tcaaatgagc
      241 gttggtcagc gggcaaagct taccatctct cccgattacg cttacggcgc aacagggcac
      301 cctggcatta tccccccaca cgcaactctc gtgtttgatg tcgagcttct taaattggaa
      361 actcggggag tacaggtgga aactattagc ccaggcgatg gccgaacctt ccctaaaaga
      421 ggccaaacct gtgtggtcca ctatacagga atgctggaag acgggaagaa ggtcgatagc
      481 agcagggacc ggaataaacc tttcaagttt atgctgggta aacaggaagt gattcggggt
      541 tgggaggagg gcgtagcaca gatgtctgtg ggccagcggg ctaaacttac tataagccct
      601 gactatgcct atggggctac cggccatcca gggattattc caccccacgc caccttggta
      661 tttgacgtgg agctcctcaa gctcgaaaca agaggggttc aggtcgagac tatttctccc
      721 ggggacggtc gcacatttcc taaacgaggt caaacatgcg tggtccatta cactggaatg
      781 ctcgaagacg gaaaaaaggt agattctagc agggatcgca acaagccatt caagttcatg
      841 ctcggaaagc aagaagtcat tcggggctgg gaagaaggcg tggcacaaat gtctgtagga
      901 cagagagcca aattgactat aagtccagat tacgcatacg gggccaccgg acatcccgga
      961 attattccac cacacgctac attggtcttt gatgtggaac tcttgaagct ggaaacacga
     1021 ggagttcaag ttgagaccat atctccaggt gatggcagaa catttcccaa gagggggcaa
     1081 acctgcgtcg tgcattacac agggatgctt gaagatggca agaaagtaga ttcctctagg
     1141 gatagaaata agccttttaa gtttatgctt ggtaaacagg aggttatcag gggatgggag
     1201 gaaggcgtcg cccagatgag cgttggtcag cgagcaaaac tcaccatcag tccagactat
     1261 gcatatggcg ccactggtca tccaggtata attcctcccc acgctacttt ggttttcgac
     1321 gttgaactgt tgaaactgga gggggccgga gctggagccg catgcaaggt actcgcccag
     1381 gagtcccaag acgtcagtgg gtctcgccag gctgtgcccc tgatcggatc tcaagccaat
     1441 tcagaagacc ggcacctcgt agaccaaatg ccagactcct ccgacaaccc agccggcctg
     1501 gctgagcaat gtccaggaat gaggaaacga cccgctgccg aggacagctc atcccaaaac
     1561 aaaagagcta accgcaccga agagaatgtg agtgacggat cacccaatct ggttatttca
     1621 gaacaaactc ctccactccc tcttagacga aggcaaacct gataagcggc cgcgtttaaa
     1681 cgtcgacggt atcgataagc ttgatatcga attccgcccc cccccctaac gttactggcc
     1741 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     1801 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     1861 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     1921 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     1981 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     2041 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     2101 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     2161 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     2221 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     2281 atggccacaa ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2341 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtctggcga gggcgagggc
     2401 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2461 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2521 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2581 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2641 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2701 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2761 aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacatcga ggacggcagc
     2821 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2881 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     2941 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3001 ctgtacaagt aatgaattaa ttaagaattc gataaaataa aagattttat ttagtctcca
     3061 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     3121 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga
     3181 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     3241 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
     3301 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     3361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     3421 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac
     3481 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     3541 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt
     3601 cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca
     3661 attagccact gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag
     3721 cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata
     3781 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3841 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3901 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3961 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4021 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4081 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4141 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4201 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4261 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4321 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4381 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4441 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4501 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4561 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4621 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4681 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4741 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4801 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4861 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4921 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4981 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5041 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5101 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5161 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5221 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5281 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5341 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5401 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5461 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5521 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5581 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5641 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5701 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5761 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5821 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5881 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     5941 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     6001 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     6061 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     6121 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     6181 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     6241 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     6301 ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg
     6361 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     6421 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     6481 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc
     6541 gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac
     6601 aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag attttattta
     6661 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     6721 taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag
     6781 gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc
     6841 tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt
     6901 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     6961 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     7021 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc
     7081 cgggtacccg tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggactcgctg
     7141 atccttggga gggtctcctc agattgattg actgcccacc tcgggggtct ttcatttgga
     7201 ggttccaccg agatttggag acccctgccc agggaccacc gacccccccg ccgggaggta
     7261 agctggccag cggtcgtttc gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct
     7321 aatgtttgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg
     7381 gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg
     7441 ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag
     7501 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
     7561 tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc
     7621 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
     7681 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     7741 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     7801 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     7861 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
     7921 gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc
     7981 cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc
     8041 ctcgatcctc cctttatcca gccctcactc cttctctagg cgccggaatt
//
