LOCUS       sequence-147760-        7189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7189)
  TITLE     toll-like receptors
REFERENCE   2  (bases 1 to 7189)
  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"
COMMENT     Sequence Label: mTLR4 flag
FEATURES             Location/Qualifiers
     source          1..7189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             932..976
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             977..1000
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     RBS             2807..2815
                     /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)"
     polyA_signal    3530..4152
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3729..3748)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4176..4196)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4181..4510
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4361..4496
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4548..4567)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4549..4567)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4581..4597)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4581..4597)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4594..4616)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4605..4621
                     /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(4629..4635,4636..4653,4654..4659))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4674..4695
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4812..4829)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4983..5571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5063..5082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5742..6533,6534..6602))
                     /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     6365..6384
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6603..6707)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6734..7162
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6821..6840)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7031..7052
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaattcac catgtctgca cttctgatcc tagctcttgt
      961 tggagctgca gttgctgact acaaagacga tgacgacaag cttgcggccg cgaattcatc
     1021 gatcctgaca ccaggaagct tgaatccctg catagaggta gttcctaata ttacctacca
     1081 atgcatggat cagaaactca gcaaagtccc tgatgacatt ccttcttcaa ccaagaacat
     1141 agatctgagc ttcaacccct tgaagatctt aaaaagctat agcttctcca atttttcaga
     1201 acttcagtgg ctggatttat ccaggtgtga aattgaaaca attgaagaca aggcatggca
     1261 tggcttacac cacctctcaa acttgatact gacaggaaac cctatccaga gtttttcccc
     1321 aggaagtttc tctggactaa caagtttaga gaatctggtg gctgtggaga caaaattggc
     1381 ctctctagaa agcttcccta ttggacagct tataacctta aagaaactca atgtggctca
     1441 caattttata cattcctgta agttacctgc atatttttcc aatctgacga acctagtaca
     1501 tgtggatctt tcttataact atattcaaac tattactgtc aacgacttac agtttctacg
     1561 tgaaaatcca caagtcaatc tctctttaga catgtctttg aacccaattg acttcattca
     1621 agaccaagcc tttcagggaa ttaagctcca tgaactgact ctaagaggta attttaatag
     1681 ctcaaatata atgaaaactt gccttcaaaa cctggctggt ttacacgtcc atcggttgat
     1741 cttgggagaa tttaaagatg aaaggaatct ggaaattttt gaaccctcta tcatggaagg
     1801 actatgtgat gtgaccattg atgagttcag gttaacatat acaaatgatt tttcagatga
     1861 tattgttaag ttccattgct tggcgaatgt ttctgcaatg tctctggcag gtgtatctat
     1921 aaaatatcta gaagatgttc ctaaacattt caaatggcaa tccttatcaa tcattagatg
     1981 tcaacttaag cagtttccaa ctctggatct accctttctt aaaagtttga ctttaactat
     2041 gaacaaaggg tctatcagtt ttaaaaaagt ggccctacca agtctcagct atctagatct
     2101 tagtagaaat gcactgagct ttagtggttg ctgttcttat tctgatttgg gaacaaacag
     2161 cctgagacac ttagacctca gcttcaatgg tgccatcatt atgagtgcca atttcatggg
     2221 tctagaagag ctgcagcacc tggattttca gcactctact ttaaaaaggg tcacagaatt
     2281 ctcagcgttc ttatcccttg aaaagctact ttaccttgac atctcttata ctaacaccaa
     2341 aattgacttc gatggtatat ttcttggctt gaccagtctc aacacattaa aaatggctgg
     2401 caattctttc aaagacaaca ccctttcaaa tgtctttgca aacacaacaa acttgacatt
     2461 cctggatctt tctaaatgtc aattggaaca aatatcttgg ggggtatttg acaccctcca
     2521 tagacttcaa ttattaaata tgagtcacaa caatctattg tttttggatt catcccatta
     2581 taaccagctg tattccctca gcactcttga ttgcagtttc aatcgcatag agacatctaa
     2641 aggaatactg caacattttc caaagagtct agccttcttc aatcttacta acaattctgt
     2701 tgcttgtata tgtgaacatc agaaattcct gcagtgggtc aaggaacaga agcagttctt
     2761 ggtgaatgtt gaacaaatga catgtgcaac acctgtagag atgaatacct ccttagtgtt
     2821 ggattttaat aattctacct gttatatgta caagacaatc atcagtgtgt cagtggtcag
     2881 tgtgattgtg gtatccactg tagcatttct gatataccac ttctattttc acctgatact
     2941 tattgctggc tgtaaaaagt acagcagagg agaaagcatc tatgatgcat ttgtgatcta
     3001 ctcgagtcag aatgaggact gggtgagaaa tgagctggta aagaatttag aagaaggagt
     3061 gccccgcttt cacctctgcc ttcactacag agactttatt cctggtgtag ccattgctgc
     3121 caacatcatc caggaaggct tccacaagag ccggaaggtt attgtggtag tgtctagaca
     3181 ctttattcag agccgttggt gtatctttga atatgagatt gctcaaacat ggcagtttct
     3241 gagcagccgc tctggcatca tcttcattgt ccttgagaag gttgagaagt ccctgctgag
     3301 gcagcaggtg gaattgtatc gccttcttag cagaaacacc tacctggaat gggaggacaa
     3361 tcctctgggg aggcacatct tctggagaag acttaaaaat gccctattgg atggaaaagc
     3421 ctcgaatcct gagcaaacag cagaggaaga acaagaaacg gcaacttgga cctgaggaga
     3481 acaaaactct ggggcctaaa cccagtctgt gtcgactcta gaggatcccg ggtggcatcc
     3541 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     3601 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     3661 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     3721 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     3781 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     3841 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     3901 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     3961 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttaaaata
     4021 actataccag caggaggacg tccagacaca gcataggcta cctggccatg cccaaccggt
     4081 gggacatttg agttgcttgc ttggcactgt cctctcatgc gttgggtcca ctcagtagat
     4141 gcctgttgaa ttgggtacgc ggccagcttg gctgtggaat gtgtgtcagt tagggtgtgg
     4201 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4261 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4321 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4381 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4441 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4501 cttttgcaaa aagctcctcg aggaactgaa aaaccagaaa gttaattccc tatagtgagt
     4561 cgtattaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4621 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4681 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4741 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4801 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4861 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4921 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4981 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5041 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5101 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5161 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5221 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5281 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5341 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5401 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5461 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5521 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5581 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5641 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5701 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5761 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5821 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5881 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5941 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6001 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6061 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6121 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6181 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6241 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6301 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6361 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6421 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6481 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6541 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6601 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6661 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6721 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     6781 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     6841 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     6901 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     6961 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     7021 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     7081 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     7141 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattt
//
