LOCUS       sequence-472886-        8453 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 8453)
  TITLE     TLR Signaling Plasmids
REFERENCE   2  (bases 1 to 8453)
  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: pcDNA3-TLR2-YFP
FEATURES             Location/Qualifiers
     source          1..8453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3271..3273,3274..3276,3277..3990)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3316..3337)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3577..3596)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3924..3945
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4006..4024)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4007..4024)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4044..4061)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4050..4274
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4320..4748
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4407..4426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4617..4638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4757..4777)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4762..5091
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4942..5077
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5004..5023
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5158..5952
                     /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(5212..5231)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5822..5841
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6126..6247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6163..6182)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6217..6236
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6296..6312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6296..6312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6309..6331)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6320..6336
                     /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(6344..6350,6351..6368,6369..6374))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6389..6410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6527..6544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6698..7286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6778..6797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7457..8248,8249..8317))
                     /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     8080..8099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8318..8422)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccatgccaca tactttgtgg atggtgtggg tcttgggggt catcatcagc
      961 ctctccaagg aagaatcctc caatcaggct tctctgtctt gtgaccgcaa tggtatctgc
     1021 aagggcagct caggatcttt aaactccatt ccctcagggc tcacagaagc tgtaaaaagc
     1081 cttgacctgt ccaacaacag gatcacctac attagcaaca gtgacctaca gaggtgtgtg
     1141 aacctccagg ctctggtgct gacatccaat ggaattaaca caatagagga agattctttt
     1201 tcttccctgg gcagtcttga acatttagac ttatcctata attacttatc taatttatcg
     1261 tcttcctggt tcaagcccct ttcttcttta acattcttaa acttactggg aaatccttac
     1321 aaaaccctag gggaaacatc tcttttttct catctcacaa aattgcaaat cctgagagtg
     1381 ggaaatatgg acaccttcac taagattcaa agaaaagatt ttgctggact taccttcctt
     1441 gaggaacttg agattgatgc ttcagatcta cagagctatg agccaaaaag tttgaagtca
     1501 attcagaacg taagtcatct gatccttcat atgaagcagc atattttact gctggagatt
     1561 tttgtagatg ttacaagttc cgtggaatgt ttggaactgc gagatactga tttggacact
     1621 ttccattttt cagaactatc cactggtgaa acaaattcat tgattaaaaa gtttacattt
     1681 agaaatgtga aaatcaccga tgaaagtttg tttcaggtta tgaaactttt gaatcagatt
     1741 tctggattgt tagaattaga gtttgatgac tgtaccctta atggagttgg taattttaga
     1801 gcatctgata atgacagagt tatagatcca ggtaaagtgg aaacgttaac aatccggagg
     1861 ctgcatattc caaggtttta cttattttat gatctgagca ctttatattc acttacagaa
     1921 agagttaaaa gaatcacagt agaaaacagt aaagtttttc tggttccttg tttactttca
     1981 caacatttaa aatcattaga atacttggat ctcagtgaaa atttgatggt tgaagaatac
     2041 ttgaaaaatt cagcctgtga ggatgcctgg ccctctctac aaactttaat tttaaggcaa
     2101 aatcatttgg catcattgga aaaaaccgga gagactttgc tcactctgaa aaacttgact
     2161 aacattgata tcagtaagaa tagttttcat tctatgcctg aaacttgtca gtggccagaa
     2221 aagatgaaat atttgaactt atccagcaca cgaatacaca gtgtaacagg ctgcattccc
     2281 aagacactgg aaattttaga tgttagcaac aacaatctca atttattttc tttgaatttg
     2341 ccgcaactca aagaacttta tatttccaga aataagttga tgactctacc agatgcctcc
     2401 ctcttaccca tgttactagt attgaaaatc agtaggaatg caataactac gttttctaag
     2461 gagcaacttg actcatttca cacactgaag actttggaag ctggtggcaa taacttcatt
     2521 tgctcctgtg aattcctctc cttcactcag gagcagcaag cactggccaa agtcttgatt
     2581 gattggccag caaattacct gtgtgactct ccatcccatg tgcgtggcca gcaggttcag
     2641 gatgtccgcc tctcggtgtc ggaatgtcac aggacagcac tggtgtctgg catgtgctgt
     2701 gctctgttcc tgctgatcct gctcacgggg gtcctgtgcc accgtttcca tggcctgtgg
     2761 tatatgaaaa tgatgtgggc ctggctccag gccaaaagga agcccaggaa agctcccagc
     2821 aggaacatct gctatgatgc atttgtttct tacagtgagc gggatgccta ctgggtggag
     2881 aaccttatgg tccaggagct ggagaacttc aatcccccct tcaagttgtg tcttcataag
     2941 cgggacttca ttcctggcaa gtggatcatt gacaatatca ttgactccat tgaaaagagc
     3001 cacaaaactg tctttgtgct ttctgaaaac tttgtgaaga gtgagtggtg caagtatgaa
     3061 ctggacttct cccatttccg tctttttgat gagaacaatg atgctgccat tctcattctt
     3121 ctggagccca ttgagaaaaa agccattccc cagcgcttct gcaagctgcg gaagataatg
     3181 aacaccaaga cctacctgga gtggcccatg gacgaggctc agcgggaagg attttgggta
     3241 aatctgagag ctgcgataaa gtccctcgag atggtgagca agggcgagga gctgttcacc
     3301 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3361 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     3421 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgaag
     3481 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3541 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3601 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3661 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     3721 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     3781 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3841 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa
     3901 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     3961 actctcggca tggacgagct gtacaagtaa tctagagggc cctattctat agtgtcacct
     4021 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4081 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     4141 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     4201 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     4261 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     4321 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4381 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4441 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4501 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4561 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4621 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4681 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4741 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4801 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4861 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4921 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4981 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5041 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     5101 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     5161 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5221 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5281 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5341 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5401 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5461 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5521 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5581 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5641 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5701 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5761 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5821 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5881 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5941 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     6001 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6061 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     6121 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6181 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6241 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6301 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6361 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6421 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6481 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6541 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6601 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6661 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6721 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6781 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6841 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6901 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6961 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7021 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7081 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7141 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7201 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7261 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7321 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7381 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7441 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7501 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7561 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7621 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7681 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7741 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7801 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7861 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7921 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7981 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8041 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8101 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8161 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8221 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8281 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8341 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8401 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
