LOCUS       sequence-263595-        8417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of  mouse TLR4 with out its cytosolic tail.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8417)
  AUTHORS   Tan Y, Zanoni I, Cullen TW, Goodman AL, Kagan JC
  TITLE     Mechanisms of Toll-like Receptor 4 Endocytosis Reveal a Common 
            Immune-Evasion Strategy Used by Pathogenic and Commensal Bacteria.
  JOURNAL   Immunity. 2015 Nov 17;43(5):909-22. doi: 
            10.1016/j.immuni.2015.10.008. Epub 2015 Nov 3.
  PUBMED    26546281
REFERENCE   2  (bases 1 to 8417)
  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.immuni.2015.10.008"; journalName: "Immunity"; date: 
            "2015-11-17- 17"; volume: "43"; issue: "5"; pages: "909-22"
COMMENT     Sequence Label: pMSCV-IRES-GFP-TLR4 delta tail
FEATURES             Location/Qualifiers
     source          1..8417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     RBS             3266..3274
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             3377..3400
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3451..4017
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3467..4019
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3611..3628)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3838..3857
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4031..4033,4034..4036,4037..4750)
                     /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(4076..4097)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4337..4356)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4684..4705
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4812..5326
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5488..5504
                     /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(5512..5518,5519..5536,5537..5542))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5557..5578
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5695..5712)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5866..6454)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5946..5965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6625..7416,7417..7485))
                     /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     7248..7267
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7486..7590)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7658..7676
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7714..7736)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7836..7855
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8049..8071
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8199..8218
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatccgcc gccaccatga tgcctccctg
     1441 gctcctggct aggactctga tcatggcact gttcttctcc tgcctgacac cgggaagctt
     1501 gaatccctgc atagaggtag ttcctaatat tacctaccaa tgcatggatc agaaactcag
     1561 caaagtccct gatgacattc cttcttcaac caagaacata gatctgagct tcaacccctt
     1621 gaagatctta aaaagctata gcttctccaa tttttcagaa cttcagtggc tggatttatc
     1681 caggtgtgaa attgaaacaa ttgaagacaa ggcatggcat ggcttacacc acctctcaaa
     1741 cttgatactg acaggaaacc ctatccagag tttttcccca ggaagtttct ctggactaac
     1801 aagtttagag aatctggtgg ctgtggagac aaaattggcc tctctagaaa gcttccctat
     1861 tggacagctt ataaccttaa agaaactcaa tgtggctcac aattttatac attcctgtaa
     1921 gttacctgca tatttttcca atctgacgaa cctagtacat gtggatcttt cttataacta
     1981 tattcaaact attactgtca atgacttaca gtttctacgt gaaaatccac aagtcaatct
     2041 ctctttagac atatctttga acccaattga cttcattcaa gaccaagcct ttcagggaat
     2101 taagctccat gaactgactc taagaggtaa ttttaatagc tcaaatataa tgaaaacttg
     2161 ccttcaaaac ctggctggtt tacacgtcca tcggttgatc ttgggagaat ttaaagatga
     2221 aaggaatctg gaaatttttg aaccctctat catggaagga ctatgtgatg tgaccattga
     2281 tgagttcagg ttaacacata caaatgattt ttcagatgat attgttaagt tccattgctt
     2341 ggcgaatgtt tctgcaatgt ctctggcagg tgtatctata aaatatctag aagatgttcc
     2401 taaacatttc aaatggcaat ccttatcaat cattagatgt caacttaagc agtttccaac
     2461 tctggatcta ccctttctta aaagtttgac tttaactatg aacaaagggt ctatcagttt
     2521 taaaaaagtg gccctgccaa gtctcagcta tctagatctt agtagaaatg cactgagctt
     2581 tagtggttgc tgttcttatt ctgatttggg aacaaacagc ctgagacact tagacctcag
     2641 cttcaatggt gccatcatta tgagtgccaa tttcatgggt ctagaagagc tgcagcacct
     2701 ggattttcag cactctactt taaaaagggt cacagaattc tcagcgttct tatcccttga
     2761 aaagctactt taccttgaca tctcttatac taacaccaaa attgacttcg atggtatatt
     2821 tcttggcttg accagtctca acacattaaa aatggctggc aattctttca aagacaacac
     2881 cctttcaaat gtctttgcaa acacaacaaa cttgacattc ctggatcttt ctaaatgtca
     2941 attggaacaa atatcttggg gggtatttga caccctccat agacttcaat tattaaatat
     3001 gagtcacaac aatctattgt ttttggattc atcccattat aaccagctgt attccctcag
     3061 cactcttgat tgcagtttca atcgcataga gacatctaaa ggaatactgc aacattttcc
     3121 aaagagtcta gccttcttca atcttactaa taattctgtt gcttgtatat gtgaacatca
     3181 gaaattcctg cagtgggtca aggaccagaa gcagttcttg gtgaatgttg aacaaatgac
     3241 atgtgcaaca cctgtagaga tgaatacctc cttagtgttg gattttaata attctacctg
     3301 ttatatgtac aagacaatca tcagtgtgtc agtgatcagt gtgattgtgg tatccactgt
     3361 agcatttctg atatacgatt acaaggatga cgacgataag tgagcggccg cgtttaaacg
     3421 tcgacggtat cgataagctt gatatcgaat tccgcccccc cccctaacgt tactggccga
     3481 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     3541 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     3601 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     3661 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     3721 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     3781 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     3841 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     3901 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     3961 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     4021 ggccacaacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     4081 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tctggcgagg gcgagggcga
     4141 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     4201 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     4261 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     4321 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     4381 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     4441 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     4501 gcagaagaac ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcagcgt
     4561 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     4621 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     4681 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     4741 gtacaagtaa tgaattaatt aagaattcga taaaataaaa gattttattt agtctccaga
     4801 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     4861 tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac
     4921 agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     4981 tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac
     5041 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     5101 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     5161 agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc
     5221 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
     5281 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatgggt aacagtttct
     5341 tgaagttgga gaacaacatt ctgagggtag gagtcgaata ttaagtaatc ctgactcaat
     5401 tagccactgt tttgaatcca catactccaa tactcctgaa atagttcatt atggacagcg
     5461 cagaaagagc tggggagaat tgtgaaattg ttatccgctc acaattccac acaacatacg
     5521 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5581 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5641 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5701 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5761 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5821 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5881 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5941 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6001 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6061 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6121 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6181 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6241 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6301 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6361 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6421 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6481 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6541 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6601 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6661 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6721 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6781 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6841 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6901 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6961 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7021 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7081 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7141 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7201 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7261 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7321 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7381 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7441 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7501 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7561 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7621 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     7681 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     7741 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     7801 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     7861 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     7921 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     7981 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     8041 agggttttcc cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag
     8101 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     8161 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     8221 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggcga
     8281 ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt gactcaacaa
     8341 tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat tttatttagt
     8401 ctccagaaaa agggggg
//
