LOCUS       sequence-446830-        8084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GMIP pLxIS (1-84) 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)
FEATURES             Location/Qualifiers
     source          1..8084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             25..539
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    701..717
                     /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(725..731,732..749,750..755))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    770..791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(908..925)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1079..1667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1159..1178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1838..2629,2630..2698))
                     /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     2461..2480
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2699..2803)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2871..2889
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2927..2949)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3049..3068
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3262..3284
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3412..3431
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             3631..4147
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    4211..4552
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             4619..5035
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4965..4987
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5003..5019
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      5048..5057
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5057..5122
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    6748..7314
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    6764..7316
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6908..6925)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7135..7154
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(7328..7330,7331..7333,7334..8047)
                     /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(7373..7394)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7634..7653)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7981..8002
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
       61 taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat
      121 caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca
      241 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
      301 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
      361 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
      421 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
      481 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg
      541 ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta
      601 atcctgactc aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc
      661 attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg ctcacaattc
      721 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
      781 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
      841 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
      901 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
      961 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1021 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1081 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1141 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1201 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1261 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1321 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1381 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1441 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1501 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1561 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1621 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1681 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1741 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1801 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1861 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     1921 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     1981 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2041 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2101 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2161 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2221 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2281 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2341 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2401 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2461 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2521 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2581 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2641 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2701 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2761 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     2821 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     2881 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2941 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     3001 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     3061 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     3121 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     3181 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     3241 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg caaggaatgg
     3301 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     3361 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     3421 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     3481 ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt
     3541 tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa
     3601 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     3661 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag
     3721 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg
     3781 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     3841 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     3901 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     3961 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     4021 actgcgtcgc ccgggtaccc gtattcccaa taaagcctct tgctgtttgc atccgaatcg
     4081 tggactcgct gatccttggg agggtctcct cagattgatt gactgcccac ctcgggggtc
     4141 tttcatttgg aggttccacc gagatttgga gacccctgcc cagggaccac cgaccccccc
     4201 gccgggaggt aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg
     4261 tgccggcatc taatgtttgc gcctgcgtct gtactagtta gctaactagc tctgtatctg
     4321 gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc
     4381 cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc
     4441 gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc
     4501 gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg
     4561 cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg
     4621 ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat
     4681 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga
     4741 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     4801 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta
     4861 catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca
     4921 ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg
     4981 ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggaat
     5041 tagatctgcc accatggact acaaggatca cgacggtgat tacaaggacc acgacatcga
     5101 ctacaaggac gacgacgaca agggcgttca agtagaaaca atatctccag gagacgggcg
     5161 tacatttcct aagcgtggcc aaacttgcgt agtccattat accggcatgc tggaggacgg
     5221 taagaaggtc gatagctcta gagaccgtaa caaacctttc aaatttatgc tggggaaaca
     5281 ggaggttatc cgtggttggg aggaaggcgt ggcccaaatg tccgttgggc agcgtgctaa
     5341 gttaaccatc tcgccagatt acgcatatgg ggctacaggc catcctggga tcattccccc
     5401 gcatgcgaca cttgtgtttg atgtagagct gttgaagttg gagacgcggg gagttcaagt
     5461 cgaaacgatt tcaccaggtg atggacgcac ctttcctaaa cggggacaaa cgtgcgtcgt
     5521 gcattacacc ggtatgctgg aagacggcaa gaaagtcgat tcaagccgcg atagaaataa
     5581 acccttcaaa ttcatgcttg gaaaacagga agtaatacgc ggatgggaag agggtgtggc
     5641 gcaaatgtct gtcgggcaaa gagctaaact gacaataagt cctgattacg cgtacggtgc
     5701 gactgggcac cctggtatca ttccgccgca cgcgacactt gtcttcgacg tcgagttact
     5761 taagcttgag acgcgggggg tgcaggtaga gactatttcg cccggtgacg gaagaacttt
     5821 cccaaagcgg ggacagacct gtgttgtcca ctacactggg atgttggagg atggtaagaa
     5881 agttgactca agtcgtgatc gcaacaaacc ctttaagttt atgcttggga agcaggaagt
     5941 catcagaggt tgggaagaag gcgtagctca gatgtcagtg ggacaacggg ccaaattaac
     6001 catcagtcca gactacgcct acggagccac cggtcaccca gggatcatac cacctcacgc
     6061 aaccttagtg ttcgatgtgg aacttttaaa gttagagacg cgcggtgtac aagtggaaac
     6121 aatcagtcca ggggatggga gaacattccc caaaagaggc cagacgtgtg tcgtgcacta
     6181 tacggggatg ttagaagatg ggaaaaaggt cgattcgtca cgcgaccgca acaaaccgtt
     6241 taaattcatg ttgggaaagc aggaggtgat tcgcggctgg gaggaaggtg tggcacagat
     6301 gagcgtgggc cagagagcaa aactgactat ctctcctgat tatgcctacg gcgcgacggg
     6361 ccaccctggc ataattccac cccacgctac cctggtattc gatgtcgagc ttctgaagct
     6421 tgagggtgct ggggctgggg ctatggactc agcggagaca gagctaaccc cagctcctga
     6481 gggcaggaag agatacagtg acatcttcca gagcctggac aacctcgaga tatcattggg
     6541 gaacgtaact tttgatcccc tggctggaga ccctgtacgc agagaagacc tggagcctga
     6601 caaagctgac acagccacag tggtgactga ggagaacagt gaagccagta gttggcggga
     6661 tctctcccca gaaggtcctg cacccctcac agagtgataa gcggccgcgt ttaaacgtcg
     6721 acggtatcga taagcttgat atcgaattcc gccccccccc ctaacgttac tggccgaagc
     6781 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     6841 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     6901 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     6961 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     7021 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     7081 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     7141 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     7201 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     7261 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc
     7321 cacaaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     7381 gctggacggc gacgtaaacg gccacaagtt cagcgtgtct ggcgagggcg agggcgatgc
     7441 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     7501 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     7561 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     7621 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     7681 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     7741 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     7801 gaagaacggc atcaaggcga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     7861 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     7921 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     7981 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     8041 caagtaatga attaattaag aattcgataa aataaaagat ttta
//
