LOCUS       sequence-265869-        7841 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 7841)
  TITLE     toll-like receptors
REFERENCE   2  (bases 1 to 7841)
  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: mTLR2
FEATURES             Location/Qualifiers
     source          1..7841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3160..3178)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3161..3178)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3198..3215)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3204..3428
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3474..3902
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3561..3580)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3771..3792
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3911..3931)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3916..4245
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4096..4231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4158..4177
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4312..5106
                     /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(4366..4385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4976..4995
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5280..5401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5317..5336)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5371..5390
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5450..5466)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5450..5466)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5463..5485)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5474..5490
                     /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(5498..5504,5505..5522,5523..5528))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5543..5564
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5681..5698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5852..6440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5932..5951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6611..7402,7403..7471))
                     /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     7234..7253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7472..7576)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7651..7670)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattct gcagatttac
      721 cgaaacctca gacaaagcgt caaatctcag aggatgctac gagctctttg gctcttctgg
      781 atcttggtgg ccataacagt cctcttcagc aaacgctgtt ctgctcagga gtctctgtca
      841 tgtgatgctt ctggggtgtg tgatggccgc tccaggtctt tcacctctat tccctccgga
      901 ctcacagcag ccatgaaaag ccttgacctg tctttcaaca agatcaccta cattggccat
      961 ggtgacctcc gagcgtgtgc gaacctccag gttctgattt tgaagtccag cagaatcaat
     1021 acaatagagg gagacgcctt ttattctctg ggcagtcttg aacatttgga tttgtctgat
     1081 aatcacctat ctagtttatc ttcctcctgg ttcgggcccc tttcctcttt gaaatactta
     1141 aacttaatgg gaaatcctta ccagacactg ggggtaacat cgctttttcc caatctcaca
     1201 aatttacaaa ccctcaggat aggaaatgta gagactttca gtgagataag gagaatagat
     1261 tttgctgggc tgacttctct caatgaactt gaaattaagg cattaagtct ccggaattat
     1321 cagtcccaaa gtctaaagtc gatccgcgac atccatcacc tgactcttca cttaagcgag
     1381 tctgctttcc tgctggagat ttttgcagat attctgagtt ctgtgagata tttagaacta
     1441 agagatacta acttggccag gttccagttt tcaccactgc ccgtagatga agtcagctca
     1501 ccgatgaaga agctggcatt ccgaggctcg gttctcactg atgaaagctt taacgagctc
     1561 ctgaagctgt tgcgttacat cttggaactg tcggaggtag agttcgacga ctgtaccctc
     1621 aatgggctcg gcgatttcaa cccctcggag tcagacgtag tgagcgagct gggtaaagta
     1681 gaaacagtca ctatccggag gttgcatatc ccccagttct atttgtttta tgacctgagt
     1741 actgtctatt ccctcctgga gaaggtgaag cgaatcacag tagagaacag caaggtcttc
     1801 ctggttccct gctcgttctc ccagcattta aaatcattag aattcttaga cctcagcgaa
     1861 aatctgatgg ttgaagaata tttgaagaac tcagcctgta agggagcctg gccttctcta
     1921 caaaccttag ttttgagcca gaatcatttg agatcaatgc aaaaaacagg agagattttg
     1981 ctgactctga aaaacctgac ctcccttgac atcagcagga acacttttca tccgatgccc
     2041 gacagctgtc agtggccaga aaagatgcgc ttcctgaatt tgtccagtac agggatccgg
     2101 gtggtaaaaa cgtgcattcc tcagacgctg gaggtgttgg atgttagtaa caacaatctt
     2161 gactcatttt ctttgttctt gcctcggctg caagagctct atatttccag aaataagctg
     2221 aaaacactcc cagatgcttc gttgttccct gtgttgctgg tcatgaaaat cagagagaat
     2281 gcagtaagta ctttctctaa agaccaactt ggttcttttc ccaaactgga gactctggaa
     2341 gcaggcgaca accactttgt ttgctcctgc gaactcctat cctttactat ggagacgcca
     2401 gctctggctc aaatcctggt tgactggcca gacagctacc tgtgtgactc tccgcctcgc
     2461 ctgcacggcc acaggcttca ggatgcccgg ccctccgtct tggaatgtca ccaggctgca
     2521 ctggtgtctg gagtctgctg tgcccttctc ctgttgatct tgctcgtagg tgccctgtgc
     2581 caccatttcc acggactgtg gtacctgaga atgatgtggg cgtggctcca ggccaagagg
     2641 aagcccaaga aagctccctg cagggacgtt tgctatgatg cctttgtttc ctacagtgag
     2701 caggattccc attgggtgga gaacctcatg gtccagcagc tggagaactc tgacccgccc
     2761 tttaagctgt gtctccacaa gcgggacttc gttccgggca aatggatcat tgacaacatc
     2821 atcgattcca tcgaaaagag ccacaaaact gtgttcgtgc tttctgagaa cttcgtacgg
     2881 agcgagtggt gcaagtacga actggacttc tcccacttca ggctctttga cgagaacaac
     2941 gacgcggcca tccttgtttt gctggagccc attgagagga aagccattcc ccagcgcttc
     3001 tgcaaactgc gcaagataat gaacaccaag acctacctgg agtggccctt ggatgaaggc
     3061 cagcaggaag tgttttgggt aaatctgaga actgcaataa agtcctaggt tctccaccca
     3121 gttcctgact tcctcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc
     3181 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3241 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3301 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3361 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3421 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3481 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3541 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3601 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3661 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3721 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3781 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3841 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3901 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3961 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4021 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4081 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4141 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4201 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4261 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4321 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4381 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4441 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4501 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4561 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4621 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4681 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4741 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4801 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4861 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4921 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4981 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5041 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5101 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5161 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5221 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5281 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5341 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5401 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5461 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5521 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5581 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5641 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5701 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5761 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5821 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5881 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5941 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6001 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6061 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6121 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6181 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6241 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6301 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6361 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6421 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6481 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6541 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6601 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6661 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6721 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6781 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6841 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6901 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6961 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7021 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7081 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7141 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7201 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7261 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7321 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7381 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7441 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7501 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7561 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7621 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     7681 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     7741 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     7801 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
