LOCUS       sequence-259500-        8283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mouse CD4 extracellular domain fused with 
            the cytosolic tail of TLR4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8283)
  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 8283)
  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-CD4-TLR4 cyto
FEATURES             Location/Qualifiers
     source          1..8283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1334..1356
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1372..1388
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             3229..3252
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     3264..3280
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3264..3280
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3494..3511)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3721..3740
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             4677..5191
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5353..5369
                     /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(5377..5383,5384..5401,5402..5407))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5422..5443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5560..5577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5731..6319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5811..5830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6490..7281,7282..7350))
                     /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     7113..7132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7351..7455)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7523..7541
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7579..7601)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7701..7720
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7914..7936
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8064..8083
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8283,1..516)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
       61 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
      121 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
      181 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
      241 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaatt agatctgccg ccaccatgtg ccgagccatc
     1441 tctcttaggc gcttgctgct gctgctgctg cagctgtcac aactcctagc tgtcactcaa
     1501 gggaagacgc tggtgctggg gaaggaaggg gaatcagcag aactgccctg cgagagttcc
     1561 cagaagaaga tcacagtctt cacctggaag ttctctgacc agaggaagat tctggggcag
     1621 catggcaaag gtgtattaat tagaggaggt tcgccttcgc agtttgatcg ttttgattcc
     1681 aaaaaagggg catgggagaa aggatcgttt cctctcatca tcaataaact taagatggaa
     1741 gactctcaga cttatatctg tgagctggag aacaggaaag aggaggtgga gttgtgggtg
     1801 ttcaaagtga ccttcagtcc gggtaccagc ctgttgcaag ggcagagcct gaccctgacc
     1861 ttggatagca actctaaggt ctctaacccc ttgacagagt gcaaacacaa aaagggtaaa
     1921 gttgtcagtg gttccaaagt tctctccatg tccaacctaa gggttcagga cagcgacttc
     1981 tggaactgca ccgtgaccct ggaccagaaa aagaactggt tcggcatgac actctcagtg
     2041 ctgggttttc agagcacagc tatcacggcc tataagagtg agggagagtc agcggagttc
     2101 tccttcccac tcaactttgc agaggaaaac gggtggggag agctgatgtg gaaggcagag
     2161 aaggattctt tcttccagcc ctggatctcc ttctccataa agaacaaaga ggtgtccgta
     2221 caaaagtcca ccaaagacct caagctccag ctgaaggaaa cgctcccact caccctcaag
     2281 ataccccagg tctcgcttca gtttgctggt tctggcaacc tgactctgac tctggacaaa
     2341 gggacactgc atcaggaagt gaacctggtg gtgatgaaag tggctcagct caacaatact
     2401 ttgacctgtg aggtgatggg acctacctct cccaagatga gactgaccct gaagcaggag
     2461 aaccaggagg ccagggtctc tgaggagcag aaagtagttc aagtggtggc ccctgagaca
     2521 gggctgtggc agtgtctact gagtgaaggt gataaggtca agatggactc caggatccag
     2581 gttttatcca gaggggtgcc atggtacaag acaatcatca gtgtgtcagt ggtcagtgtg
     2641 attgtggtat ccactgtagc atttctgata taccacttct attttcacct gatacttatt
     2701 gctggctgta aaaagtacag cagaggagaa agcatctatg atgcatttgt gatctactcg
     2761 agtcagaatg aggactgggt gagaaatgag ctggtaaaga atttagaaga aggagtgccc
     2821 cgctttcacc tctgccttca ctacagagac tttattcctg gtgtagccat tgctgccaac
     2881 atcatccagg aaggcttcca caagagccgg aaggttattg tggtagtgtc tagacacttt
     2941 attcagagcc gttggtgtat ctttgaatat gagattgctc aaacatggca gtttctgagc
     3001 agccgctctg gcatcatctt cattgtcctt gagaaggttg agaagtccct gctgaggcag
     3061 caggtggaat tgtatcgcct tcttagcaga aacacctacc tggaatggga ggacaatcct
     3121 ctggggaggc acatcttctg gagaagactt aaaaatgccc tattggatgg aaaagcctcg
     3181 aatcctgagc aaacagcaga ggaagaacaa gaaacggcaa cttggaccga ttacaaggat
     3241 gacgacgata agtgagcggc cgctctagaa ctagtggatc ccccgggctg catctagggc
     3301 ggccaattcc gcccctctcc cccccccccc tctccctccc ccccccctaa cgttactggc
     3361 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     3421 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     3481 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     3541 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     3601 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     3661 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     3721 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     3781 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     3841 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgccac
     3901 catgagcttt ccatgtaaat ttgtagccag cttccttctg attttcaatg tttcttccaa
     3961 aggtgcagtc tccaaagaga ttacgaatgc cttggaaacc tggggtgcct tgggtcagga
     4021 catcaacttg gacattccta gttttcaaat gagtgatgat attgacgata taaaatggga
     4081 aaaaacttca gacaagaaaa agattgcaca attcagaaaa gagaaagaga ctttcaagga
     4141 aaaagataca tataagctat ttaaaaatgg aactctgaaa attaagcatc tgaagaccga
     4201 tgatcaggat atctacaagg tatcaatata tgatacaaaa ggaaaaaatg tgttggaaaa
     4261 aatatttgat ttgaagattc aagagagggt ctcaaaacca aaaatctcct ggacttgtat
     4321 caacacaacc ctgacctgtg aggtaatgaa tggaactgac cccgaattaa acctgtatca
     4381 agatgggaaa catctaaaac tttctcagag ggtcatcaca cacaagtgga ccaccagcct
     4441 gagtgcaaaa ttcaagtgca cagcagggaa caaagtcagc aaggaatcca gtgtcgagcc
     4501 tgtcagctgt ccagagaaag gtctggacat ctatctcatc attggcatat gtggaggagg
     4561 cagcctcttg atggtctttg tggcactgct cgttttctat atcaccaaaa ggaaaaaaca
     4621 gaggtaggaa ttcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     4681 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     4741 aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa
     4801 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     4861 gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca
     4921 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     4981 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     5041 actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtgta tccaataaac
     5101 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     5161 gattgactac ccgtcagcgg gggtctttca tgggtaacag tttcttgaag ttggagaaca
     5221 acattctgag ggtaggagtc gaatattaag taatcctgac tcaattagcc actgttttga
     5281 atccacatac tccaatactc ctgaaatagt tcattatgga cagcgcagaa agagctgggg
     5341 agaattgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5401 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5461 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5521 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5581 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5641 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5701 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5761 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5821 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5881 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5941 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6001 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6061 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6121 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6181 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6241 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6301 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6361 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6421 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6481 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6541 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6601 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6661 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6721 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6781 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6841 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6901 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6961 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7021 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7081 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7141 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7201 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7261 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7321 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7381 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7441 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     7501 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     7561 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     7621 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     7681 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     7741 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     7801 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     7861 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     7921 acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     7981 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     8041 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     8101 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt
     8161 aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag
     8221 cctatagagt acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg
     8281 gga
//
