LOCUS       sequence-142442-        7949 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 7949)
  TITLE     toll-like receptors
REFERENCE   2  (bases 1 to 7949)
  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: hTLR4
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    270..391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(281..300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     335..354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(565..1359)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(676..695)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1286..1305
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1426..1755)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1440..1575
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1494..1513)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1740..1760
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1769..2197)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1879..1900)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2091..2110
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2243..2467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2456..2473
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2493..2511
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2493..2510
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5131..5150)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5132..5150)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5195..5398)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5224..5244)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5399..5778
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5950..5969
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6044..6148
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6149..6217,6218..7009)
                     /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     complement(6367..6386)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7180..7768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7669..7688
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7922..7939
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
      241 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
      301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      361 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
      421 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
      481 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
      541 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
      601 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
      661 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
      721 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
      781 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
      841 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
      901 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
      961 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     1021 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     1081 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     1141 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     1201 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     1261 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     1321 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     1381 ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag
     1441 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc
     1501 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     1561 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     1621 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     1681 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     1741 ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca
     1801 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     1861 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     1921 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     1981 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2041 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2101 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2161 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag
     2221 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
     2281 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     2341 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     2401 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     2461 ggcacagtcg aggctgatca gcgagctcta gcatttaggt gacactatag aatagggccc
     2521 tctagatgca tgctcgagaa gaaatgcctc aggaggtttt tatttttcct cttcagatag
     2581 atgttgcttc ctgccaattg catcctgtac ccactgttcc ttctggattc catgatttac
     2641 catccagcag ggcttttctg agtcgtctcc agaagatgtg ccgccccagg acactgtcct
     2701 cccactccag gtaagtgttc ctgctgagaa ggcggtacag ctccacctgc tgcctgagca
     2761 gggtcttctc caccttctgc aggacaatga agatgatacc agcacgactg ctcagaaact
     2821 gccaggtctg agcaatctca tattcaaaga tacaccagcg gctctggatg aagtgctggg
     2881 acaccacaac aatcaccttt cggcttttat ggaaaccttc atggatgatg ttggcagcaa
     2941 tggccacacc gggaataaag tctctgtagt gaaggcagag ctgaaatgga ggcacccctt
     3001 cttctaaatt ctttactagc tcattcctta cccagtcctc atcctggctt gagtagataa
     3061 caaaggcatc atagatgttt tcacctctac catactttat gcagccagca agaagcatca
     3121 ggtgaaaata gaacttatag accagaactg ctacaacaga tactacaagc acactgagga
     3181 ccgacacacc aatgatggtc ttattcatct gacaggtgat attcaaactc agcacaggca
     3241 tgccctgctt atctgaaggt gttgcacatt ccattcgttc aacttccacc aagagctgcc
     3301 tctggtcctt gatccattgc aggaaactct ggtgttcaca agtacaagca aagtcattct
     3361 gagtaagatt taagaaagct agactacttg gaaaatgctg tagttcctgt tttttggaag
     3421 tcattatgtg attgagactg taatcaagaa cctggaggga gttcagacac ttataaggaa
     3481 acgtatccaa tgaaaagaag ttgttgtggc tcatatttag tacctgaaga ctggagagtg
     3541 agttaaatgc tgttggagac aactgctcca gttgacactg agagaggtcc aggaaggtca
     3601 agtttctcag ctctgtgaag atatctggaa ggaagttttc ctggaaagaa ttgccagcca
     3661 ttttcaagac ttcgagactg gacaagccat tgaagatgcc attgaaagca actctggtgt
     3721 gagtatgaga aatgtcaagg taaatgaggt ttctgagtga taggaatact gaaaactcac
     3781 tcatttgttt caaattggaa tgctggaaat ccagatgttc tagttgttct aagcccaaga
     3841 agtttgaact catggtaata acaccattga agctcagatc taaatacttt aggctggttg
     3901 tcccaaaatc actttgagaa cagcaacctt tgaaactcaa gccatttcta ctgagatcta
     3961 gaaactcaag gcttggtaga tcaacttctg aaaaagcatt cccacctttg ttggaagtga
     4021 aagtaagcct tttgagagat ttgagtttca atgtgggaaa ctgtccaaat ttacagttaa
     4081 ctaattctaa atgttgccat ccgaaattat aagaaaagtc ttttaccctt tcaatagtca
     4141 cactcaccag ggaaaatgaa gaaacatttg tcaaacaatt aaataagtca ataatatcat
     4201 cgaggtagta gtctaagtat gctaatcgga attcttcaat ggtcaaattg cacaggccct
     4261 ctagagcaga tttgtcaaac ttttccaagt ttccttcatt tctaaattct cccagaacca
     4321 aacgatggac ttctaaacca gccagacctt gaatacaagt tttcattaca tttaaactat
     4381 caaaattatt tcttaaagtc agcttatgaa gcctaatttc tttaaatgca cctggttgga
     4441 taaagttcat agggttcagg gacaggtcta aagagagatt gagtaggggc atttgatgta
     4501 gaacccgcaa gtctgtgcaa taaatacttt gaatcttgtt gctggaaagg tccaagtgct
     4561 ctagattggt cagattagaa aaatactcag gtaatttgaa agattggata agattgtgag
     4621 ccacattaag ttctttcaaa gttttgagat gtccaatggg gaagttctct agagatgcta
     4681 gatttgtctc cacagccacc agcttctgta aacttgatag tccagaaaag gctcccaggg
     4741 ctaaactctg gatggggttt cctgtcaata ttaaggtaga gaggtggctt aggctctgat
     4801 atgccccatc ttcaattgtc tggatttcac acctggataa atccagcacc tgcagttctg
     4861 ggaaactgaa gaagctatag ctgcctaaat gcctcagggg attaaagctc aggtccaggt
     4921 tcttggttga gaaggggagg ttgtcgggga ttttgtagaa attcagctcc atgcattgat
     4981 aagtaatatt aggaaccacc tccacgcagg gctcccagct ttctggtctc acgcaggaga
     5041 ggaaggccat ggctgggatc agagtcccag ccaggcgcga ggcagacatc atcctggcat
     5101 catcctcact ggttttgtaa gcttgggtct ccctatagtg agtcgtatta atttcgataa
     5161 gccagtaagc agtgggttct ctagttagcc agagagctct gcttatatag acctcccacc
     5221 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
     5281 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
     5341 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca
     5401 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     5461 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     5521 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     5581 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     5641 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     5701 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     5761 tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac atcgcgaagc agcgcaaaac
     5821 gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt gttgtagctt
     5881 aaattttgct cgcgcactac tcagcgacct ccaacacaca agcagggagc agatactggc
     5941 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagggga tcgggagatc
     6001 tcccgatccg tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6061 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6121 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6181 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6241 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     6301 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6361 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     6421 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     6481 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     6541 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     6601 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     6661 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     6721 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6781 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6841 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6901 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6961 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7021 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7081 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7141 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7201 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7261 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7321 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7381 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7441 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7501 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7561 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7621 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7681 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7741 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7801 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     7861 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     7921 cagcgagtca gtgagcgagg aagcggaag
//
