LOCUS       sequence-83361-e        7143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of mutant TLR4 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7143)
  AUTHORS   Kuo WT, Lee TC, Yang HY, Chen CY, Au YC, Lu YZ, Wu LL, Wei SC, Ni 
            YH, Lin BR, Chen Y, Tsai YH, Kung JT, Sheu F, Lin LW, Yu LC
  TITLE     LPS receptor subunits have antagonistic roles in epithelial 
            apoptosis and colonic carcinogenesis.
  JOURNAL   Cell Death Differ. 2015 Jan 30. doi: 10.1038/cdd.2014.240.
  PUBMED    25633197
REFERENCE   2  (bases 1 to 7143)
  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"; journalName: "Cell Death 
            Differ. 2015 Jan 30. doi: 10.1038/cdd.2014.240."
COMMENT     Sequence Label: pMyc-CMV1-huTLR4mut-A896G
FEATURES             Location/Qualifiers
     source          1..7143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..150
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     142..159
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     143..159
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        312..690
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        691..894
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     845..865
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             925..969
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             970..999
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3482..4106
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4128..4148)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4133..4463
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4314..4449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4501..4520)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4502..4520)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4534..4550)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4534..4550)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4547..4569)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4558..4574
                     /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(4582..4588,4589..4606,4607..4612))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4627..4648
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4765..4782)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4936..5524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5016..5035)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5695..6486,6487..6555))
                     /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     6318..6337
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6556..6660)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6687..7143
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6774..6793)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6984..7005
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccca
      121 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctgat ctaatcaata
      181 ttggccatta gccatattat tcattggtta tatagcataa atcaatattg gctattggcc
      241 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt
      301 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      361 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      421 cgaccgccca gcgacccccg cccgttgacg tcaatagtga cgtatgttcc catagtaacg
      481 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      541 gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa
      601 tggcccgcct agcattatgc ccagtacatg accttacggg agtttcctac ttggcagtac
      661 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg
      721 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      781 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg
      841 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      901 gtgaaccgtc agaattaatt caccatgtct gcacttctga tcctagctct tgttggagct
      961 gcagttgctg aacaaaaact catctcagaa gaggatctgc ttgcggccgc ggaaagctgg
     1021 gagccctgcg tggaggtggt tcctaatatt acttatcaat gcatggagct gaatttctac
     1081 aaaatccccg acaacctccc cttctcaacc aagaacctgg acctgagctt taatcccctg
     1141 aggcatttag gcagctatag cttcttcagt ttcccagaac tgcaggtgct ggatttatcc
     1201 aggtgtgaaa tccagacaat tgaagatggg gcatatcaga gcctaagcca cctctctacc
     1261 ttaatattga caggaaaccc catccagagt ttagccctgg gagccttttc tggactatca
     1321 agtttacaga agctggtggc tgtggagaca aatctagcat ctctagagaa cttccccatt
     1381 ggacatctca aaactttgaa agaacttaat gtggctcaca atcttatcca atctttcaaa
     1441 ttacctgagt atttttctaa tctgaccaat ctagagcact tggacctttc cagcaacaag
     1501 attcaaagta tttattgcac agacttgcgg gttctacatc aaatgcccct actcaatctc
     1561 tctttagacc tgtccctgaa ccctatgaac tttatccaac caggtgcatt taaagaaatt
     1621 aggcttcata agctgacttt aagaaataat tttgatagtt taaatgtaat gaaaacttgt
     1681 attcaaggtc tggctggttt agaagtccat cgtttggttc tgggagaatt tagaaatgaa
     1741 ggaaacttgg aaaagtttga caaatctgct ctagagggcc tgtgcaattt gaccattgaa
     1801 gaattccgat tagcatactt agactactac ctcgatggta ttattgactt atttaattgt
     1861 ttgacaaatg tttcttcatt ttccctggtg agtgtgacta ttgaaagggt aaaagacttt
     1921 tcttataatt tcggatggca acatttagaa ttagttaact gtaaatttgg acagtttccc
     1981 acattgaaac tcaaatctct caaaaggctt actttcactt ccaacaaagg tgggaatgct
     2041 ttttcagaag ttgatctacc aagccttgag tttctagatc tcagtagaaa tggcttgagt
     2101 ttcaaaggtt gctgttctca aagtgatttt gggacaacca gcctaaagta tttagatctg
     2161 agcttcaatg gtgttattac catgagttca aacttcttgg gcttagaaca actagaacat
     2221 ctggatttcc agcattccaa tttgaaacaa atgagtgagt tttcagtatt cctatcactc
     2281 agaaacctca tttaccttga catttctcat actcacacca gagttgcttt caatggcatc
     2341 ttcaatggct tgtccagtct cgaagtcttg aaaatggctg gcaattcttt ccaggaaaac
     2401 ttccttccag atatcttcac agagctgaga aacttgacct tcctggacct ctctcagtgt
     2461 caactggagc agttgtctcc aacagcattt aactcactct ccagtcttca ggtactaaat
     2521 atgagccaca acaacttctt ttcattggat acgtttcctt ataagtgtct gaactccctc
     2581 caggttcttg attacagtct caatcacata atgacttcca aaaaacagga actacagcat
     2641 tttccaagta gtctagcttt cttaaatctt actcagaatg actttgcttg tacttgtgaa
     2701 caccagagtt tcctgcaatg gatcaaggac cagaggcagc tcttggtgga agttgaacga
     2761 atggaatgtg caacaccttc agataagcag ggcatgcctg tgctgagttt gaatatcacc
     2821 tgtcagatga ataagaccat cattggtgtg tcggtcctca gtgtgcttgt agtatctgtt
     2881 gtagcagttc tggtctataa gttctatttt cacctgatgc ttcttgctgg ctgcataaag
     2941 tatggtagag gtgaaaacat ctatgatgcc tttgttatct actcaagcca ggatgaggac
     3001 tgggtaagga atgagctagt aaagaattta gaagaagggg tgcctccatt tcagctctgc
     3061 cttcactaca gagactttat tcccggtgtg gccattgctg ccaacatcat ccatgaaggt
     3121 ttccataaaa gccgaaaggt gattgttgtg gtgtcccagc acttcatcca gagccgctgg
     3181 tgtatctttg aatatgagat tgctcagacc tggcagtttc tgagcagtcg tgctggtatc
     3241 atcttcattg tcctgcagaa ggtggagaag accctgctca ggcagcaggt ggagctgtac
     3301 cgccttctca gcaggaacac ttacctggag tgggaggaca gtgtcctggg gcggcacatc
     3361 ttctggagac gactcagaaa agccctgctg gatggtaaat catggaatcc agaaggaaca
     3421 gtgggtacag gatgcaattg gcaggaagca acatctatct gagtcgactc tagaggatcc
     3481 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccttggaag ttgccactcc
     3541 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     3601 ctctataata ttatggggtg gaggggggtg gtatggagca aggggcccaa gttgggaaga
     3661 caacctgtag ggcctgcggg gtctattcgg gaaccaagct ggagtgcagt ggcacaatct
     3721 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag
     3781 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     3841 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     3901 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     3961 ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag gctacctgcc
     4021 atggcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg
     4081 tccactcagt agatgcctgt tgaattgggt acgcggccag cttctgtgga atgtgtgtca
     4141 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4201 caattagtca gcaaccaggt gtggaaaagt ccccaggctc cccagcaggc agaagtatgc
     4261 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4321 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4381 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4441 ttggaggcct aggcttttgc aaaaagctcc tcgaggaact gaaaaaccag aaagttaatt
     4501 ccctatagtg agtcgtatta aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     4561 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4621 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4681 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4741 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4801 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4861 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4921 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4981 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5041 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5101 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     5161 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5221 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5281 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5341 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5401 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5461 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5521 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5581 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5641 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5701 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5761 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5821 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5881 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5941 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6001 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6061 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6121 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6181 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6241 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6301 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6361 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6421 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6481 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6541 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6601 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6661 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     6721 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     6781 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     6841 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     6901 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     6961 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     7021 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     7081 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa acgtttacaa
     7141 ttt
//
