LOCUS       sequence-259547-        7554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mouse CD14 mutant ""4R"" deficient in LPS 
            binding.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7554)
  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 7554)
  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-hCD2-CD14 (4R)
FEATURES             Location/Qualifiers
     source          1..7554
                     /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"
     regulatory      1420..1429
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2535..2551
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2535..2551
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2765..2782)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2992..3011
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             3948..4462
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4624..4640
                     /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(4648..4654,4655..4672,4673..4678))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4693..4714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4831..4848)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5002..5590)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5082..5101)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5761..6552,6553..6621))
                     /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     6384..6403
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6622..6726)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6794..6812
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6850..6872)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6972..6991
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7185..7207
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7335..7354
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7554,1..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 agatccgccg ccaccatgga gcgtgtgctt
     1441 ggcttgttgc tgttgcttct ggtgcacgcc tctcccgccc caccagagcc ctgcgagcta
     1501 gccgcggcag cttgttcctg caacttctca gatgcggcgg cagcttggtc cagcgctttc
     1561 aattgtttgg gggcggcagc tgcggcattg tacggcggag gccgcagcct ggaatacctt
     1621 ctaaagcgtg tggacacgga agcagctgcg gcggcgttca ctgatattat caagtctctg
     1681 tccttaaagc ggcttacggt gcgggccgcg cggattccta gtcggattct attcggagcc
     1741 ctgcgtgtgc tcgggatttc cggcctccag gaactgactc ttgaaaatct cgaggtaacc
     1801 ggcaccgcgc cgccaccgct tctggaagcc accggacccg atctcaacat cttgaacctc
     1861 cgcaacgtgt cgtgggcaac aagggatgcc tggctcgcag aactgcagca gtggctaaag
     1921 cctggactca aggtactgag tattgcccaa gcacactcac tcaacttttc ctgcgaacag
     1981 gtccgcgtct tccctgccct ctccacctta gacctgtctg acaatcctga attgggcgag
     2041 agaggactga tctcagccct ctgtcccctc aagttcccga ccctccaagt tttagcgctg
     2101 cgtaacgcgg ggatggagac gcccagcggc gtgtgctctg cgctggccgc agcaagggta
     2161 cagctgcaag gactagacct tagtcacaat tcactgcggg atgctgcagg cgctccgagt
     2221 tgtgactggc ccagtcagct aaactcgctc aatctgtctt tcactgggct gaagcaggta
     2281 cctaaagggc tgccagccaa gctcagcgtg ctggatctca gttacaacag gctggatagg
     2341 aaccctagcc cagatgagct gccccaagtg gggaacctgt cacttaaagg aaatcccttt
     2401 ttggactctg aatcccactc ggagaagttt aactctggcg tagtcaccgc cggagctcca
     2461 tcatcccaag cagtggcctt gtcaggaact ctggctttgc tcctaggaga tcgcctcttt
     2521 gtttaagcgg ccgctctaga actagtggat cccccgggct gcatctaggg cggccaattc
     2581 cgcccctctc cccccccccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
     2641 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     2701 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     2761 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     2821 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     2881 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     2941 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     3001 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     3061 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     3121 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgcca ccatgagctt
     3181 tccatgtaaa tttgtagcca gcttccttct gattttcaat gtttcttcca aaggtgcagt
     3241 ctccaaagag attacgaatg ccttggaaac ctggggtgcc ttgggtcagg acatcaactt
     3301 ggacattcct agttttcaaa tgagtgatga tattgacgat ataaaatggg aaaaaacttc
     3361 agacaagaaa aagattgcac aattcagaaa agagaaagag actttcaagg aaaaagatac
     3421 atataagcta tttaaaaatg gaactctgaa aattaagcat ctgaagaccg atgatcagga
     3481 tatctacaag gtatcaatat atgatacaaa aggaaaaaat gtgttggaaa aaatatttga
     3541 tttgaagatt caagagaggg tctcaaaacc aaaaatctcc tggacttgta tcaacacaac
     3601 cctgacctgt gaggtaatga atggaactga ccccgaatta aacctgtatc aagatgggaa
     3661 acatctaaaa ctttctcaga gggtcatcac acacaagtgg accaccagcc tgagtgcaaa
     3721 attcaagtgc acagcaggga acaaagtcag caaggaatcc agtgtcgagc ctgtcagctg
     3781 tccagagaaa ggtctggaca tctatctcat cattggcata tgtggaggag gcagcctctt
     3841 gatggtcttt gtggcactgc tcgttttcta tatcaccaaa aggaaaaaac agaggtagga
     3901 attcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     3961 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata
     4021 actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca
     4081 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     4141 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     4201 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     4261 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     4321 gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     4381 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     4441 cccgtcagcg ggggtctttc atgggtaaca gtttcttgaa gttggagaac aacattctga
     4501 gggtaggagt cgaatattaa gtaatcctga ctcaattagc cactgttttg aatccacata
     4561 ctccaatact cctgaaatag ttcattatgg acagcgcaga aagagctggg gagaattgtg
     4621 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4681 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4741 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4801 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4861 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4921 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4981 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5041 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5101 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5161 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5221 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5281 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5341 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5401 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5461 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5521 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5581 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5641 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5701 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5761 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5821 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5881 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5941 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6001 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6061 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6121 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6181 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6241 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6301 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6361 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6421 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6481 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6541 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6601 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6661 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6721 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     6781 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     6841 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     6901 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     6961 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     7021 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     7081 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     7141 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     7201 taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     7261 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     7321 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     7381 gtgatgccgg ccacgatgcg tccggcgtag aggcgattag tccaatttgt taaagacagg
     7441 atatcagtgg tccaggctct agttttgact caacaatatc accagctgaa gcctatagag
     7501 tacgagccat agataaaata aaagatttta tttagtctcc agaaaaaggg ggga
//
