LOCUS       sequence-438518-        8084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GMIP pLxIS mutant in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8084)
  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 8084)
  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-GMIP(pLxIS) S31A
FEATURES             Location/Qualifiers
     source          1..8084
                     /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..81
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    1707..2273
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1723..2275
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1867..1884)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2094..2113
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2287..2289,2290..2292,2293..3006)
                     /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(2332..2353)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2593..2612)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2940..2961
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3068..3582
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3744..3760
                     /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(3768..3774,3775..3792,3793..3798))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3813..3834
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3951..3968)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4122..4710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4202..4221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4881..5672,5673..5741))
                     /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     5504..5523
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5742..5846)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5914..5932
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5970..5992)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6092..6111
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6305..6327
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6455..6474
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6674..7190
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7254..7595
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7662..8078
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8008..8030
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8046..8062
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 agatctgcca ccatggacta caaggatcac gacggtgatt acaaggacca cgacatcgac
       61 tacaaggacg acgacgacaa gggcgttcaa gtagaaacaa tatctccagg agacgggcgt
      121 acatttccta agcgtggcca aacttgcgta gtccattata ccggcatgct ggaggacggt
      181 aagaaggtcg atagctctag agaccgtaac aaacctttca aatttatgct ggggaaacag
      241 gaggttatcc gtggttggga ggaaggcgtg gcccaaatgt ccgttgggca gcgtgctaag
      301 ttaaccatct cgccagatta cgcatatggg gctacaggcc atcctgggat cattcccccg
      361 catgcgacac ttgtgtttga tgtagagctg ttgaagttgg agacgcgggg agttcaagtc
      421 gaaacgattt caccaggtga tggacgcacc tttcctaaac ggggacaaac gtgcgtcgtg
      481 cattacaccg gtatgctgga agacggcaag aaagtcgatt caagccgcga tagaaataaa
      541 cccttcaaat tcatgcttgg aaaacaggaa gtaatacgcg gatgggaaga gggtgtggcg
      601 caaatgtctg tcgggcaaag agctaaactg acaataagtc ctgattacgc gtacggtgcg
      661 actgggcacc ctggtatcat tccgccgcac gcgacacttg tcttcgacgt cgagttactt
      721 aagcttgaga cgcggggggt gcaggtagag actatttcgc ccggtgacgg aagaactttc
      781 ccaaagcggg gacagacctg tgttgtccac tacactggga tgttggagga tggtaagaaa
      841 gttgactcaa gtcgtgatcg caacaaaccc tttaagttta tgcttgggaa gcaggaagtc
      901 atcagaggtt gggaagaagg cgtagctcag atgtcagtgg gacaacgggc caaattaacc
      961 atcagtccag actacgccta cggagccacc ggtcacccag ggatcatacc acctcacgca
     1021 accttagtgt tcgatgtgga acttttaaag ttagagacgc gcggtgtaca agtggaaaca
     1081 atcagtccag gggatgggag aacattcccc aaaagaggcc agacgtgtgt cgtgcactat
     1141 acggggatgt tagaagatgg gaaaaaggtc gattcgtcac gcgaccgcaa caaaccgttt
     1201 aaattcatgt tgggaaagca ggaggtgatt cgcggctggg aggaaggtgt ggcacagatg
     1261 agcgtgggcc agagagcaaa actgactatc tctcctgatt atgcctacgg cgcgacgggc
     1321 caccctggca taattccacc ccacgctacc ctggtattcg atgtcgagct tctgaagctt
     1381 gagggtgctg gggctggggc tatggactca gcggagacag agctaacccc agctcctgag
     1441 ggcaggaaga gatacagtga catcttccag agcctggaca acctcgagat agccttgggg
     1501 aacgtaactt ttgatcccct ggctggagac cctgtacgca gagaagacct ggagcctgac
     1561 aaagctgaca cagccacagt ggtgactgag gagaacagtg aagccagtag ttggcgggat
     1621 ctctccccag aaggtcctgc acccctcaca gagtgataag cggccgcgtt taaacgtcga
     1681 cggtatcgat aagcttgata tcgaattccg cccccccccc taacgttact ggccgaagcc
     1741 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     1801 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     1861 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     1921 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     1981 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     2041 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     2101 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     2161 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     2221 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
     2281 acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2341 ctggacggcg acgtaaacgg ccacaagttc agcgtgtctg gcgagggcga gggcgatgcc
     2401 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2461 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     2521 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2581 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2641 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2701 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     2761 aagaacggca tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     2821 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2881 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     2941 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3001 aagtaatgaa ttaattaaga attcgataaa ataaaagatt ttatttagtc tccagaaaaa
     3061 ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg
     3121 caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt aggaacagag
     3181 agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     3241 ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc
     3301 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     3361 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     3421 cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt
     3481 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     3541 ctcctctgag tgattgacta cccgtcagcg ggggtctttc atgggtaaca gtttcttgaa
     3601 gttggagaac aacattctga gggtaggagt cgaatattaa gtaatcctga ctcaattagc
     3661 cactgttttg aatccacata ctccaatact cctgaaatag ttcattatgg acagcgcaga
     3721 aagagctggg gagaattgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3781 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3841 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3901 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3961 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4021 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4081 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4141 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4201 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4261 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4321 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4381 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4441 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4501 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4561 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4621 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4681 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4741 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4801 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4861 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4921 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4981 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5041 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5101 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5161 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5221 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5281 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5341 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5401 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5461 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5521 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5581 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5641 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5701 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5761 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5821 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5881 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5941 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     6001 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     6061 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     6121 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     6181 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     6241 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     6301 ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg
     6361 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     6421 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     6481 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggcgattag
     6541 tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc
     6601 accagctgaa gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc
     6661 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
     6721 cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg
     6781 aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     6841 ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag
     6901 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     6961 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     7021 aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta
     7081 cccgtattcc caataaagcc tcttgctgtt tgcatccgaa tcgtggactc gctgatcctt
     7141 gggagggtct cctcagattg attgactgcc cacctcgggg gtctttcatt tggaggttcc
     7201 accgagattt ggagacccct gcccagggac caccgacccc cccgccggga ggtaagctgg
     7261 ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc atctaatgtt
     7321 tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
     7381 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
     7441 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
     7501 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
     7561 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
     7621 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
     7681 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     7741 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     7801 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     7861 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     7921 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     7981 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     8041 cctcccttta tccagccctc actccttctc taggcgccgg aatt
//
