LOCUS       sequence-339915-        8618 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 8618)
  TITLE     TLR Signaling Plasmids
REFERENCE   2  (bases 1 to 8618)
  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-TLR4-YFP
FEATURES             Location/Qualifiers
     source          1..8618
                     /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(3436..3438,3439..3441,3442..4155)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3481..3502)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3742..3761)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4089..4110
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4171..4189)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4172..4189)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4209..4226)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4215..4439
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4485..4913
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4572..4591)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4782..4803
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4922..4942)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4927..5256
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5107..5242
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5169..5188
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5323..6117
                     /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(5377..5396)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5987..6006
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6291..6412
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6328..6347)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6382..6401
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6461..6477)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6461..6477)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6474..6496)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6485..6501
                     /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(6509..6515,6516..6533,6534..6539))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6554..6575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6692..6709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6863..7451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6943..6962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7622..8413,8414..8482))
                     /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     8245..8264
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8483..8587)
                     /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 ccatgatgtc tgcctcgcgc ctggctggga ctctgatccc agccatggcc
      961 ttcctctcct gcgtgagacc agaaagctgg gagccctgcg tggaggtggt tcctaatatt
     1021 acttatcaat gcatggagct gaatttctac aaaatccccg acaacctccc cttctcaacc
     1081 aagaacctgg acctgagctt taatcccctg aggcatttag gcagctatag cttcttcagt
     1141 ttcccagaac tgcaggtgct ggatttatcc aggtgtgaaa tccagacaat tgaagatggg
     1201 gcatatcaga gcctaagcca cctctctacc ttaatattga caggaaaccc catccagagt
     1261 ttagccctgg gagccttttc tggactatca agtttacaga agctggtggc tgtggagaca
     1321 aatctagcat ctctagagaa cttccccatt ggacatctca aaactttgaa agaacttaat
     1381 gtggctcaca atcttatcca atctttcaaa ttacctgagt atttttctaa tctgaccaat
     1441 ctagagcact tggacctttc cagcaacaag attcaaagta tttattgcac agacttgcgg
     1501 gttctacatc aaatgcccct actcaatctc tctttagacc tgtccctgaa ccctatgaac
     1561 tttatccaac caggtgcatt taaagaaatt aggcttcata agctgacttt aagaaataat
     1621 tttgatagtt taaatgtaat gaaaacttgt attcaaggtc tggctggttt agaagtccat
     1681 cgtttggttc tgggagaatt tagaaatgaa ggaaacttgg aaaagtttga caaatctgct
     1741 ctagagggcc tgtgcaattt gaccattgaa gaattccgat tagcatactt agactactac
     1801 ctcgatgata ttattgactt atttaattgt ttgacaaatg tttcttcatt ttccctggtg
     1861 agtgtgacta ttgaaagggt aaaagacttt tcttataatt tcggatggca acatttagaa
     1921 ttagttaact gtaaatttgg acagtttccc acattgaaac tcaaatctct caaaaggctt
     1981 actttcactt ccaacaaagg tgggaatgct ttttcagaag ttgatctacc aagccttgag
     2041 tttctagatc tcagtagaaa tggcttgagt ttcaaaggtt gctgttctca aagtgatttt
     2101 gggacaacca gcctaaagta tttagatctg agcttcaatg gtgttattac catgagttca
     2161 aacttcttgg gcttagaaca actagaacat ctggatttcc agcattccaa tttgaaacaa
     2221 atgagtgagt tttcagtatt cctatcactc agaaacctca tttaccttga catttctcat
     2281 actcacacca gagttgcttt caatggcatc ttcaatggct tgtccagtct cgaagtcttg
     2341 aaaatggctg gcaattcttt ccaggaaaac ttccttccag atatcttcac agagctgaga
     2401 aacttgacct tcctggacct ctctcagtgt caactggagc agttgtctcc aacagcattt
     2461 aactcactct ccagtcttca ggtactaaat atgagccaca acaacttctt ttcattggat
     2521 acgtttcctt ataagtgtct gaactccctc caggttcttg attacagtct caatcacata
     2581 atgacttcca aaaaacagga actacagcat tttccaagta gtctagcttt cttaaatctt
     2641 actcagaatg actttgcttg tacttgtgaa caccagagtt tcctgcaatg gatcaaggac
     2701 cagaggcagc tcttggtgga agttgaacga atggaatgtg caacaccttc agataagcag
     2761 ggcatgcctg tgctgagttt gaatatcacc tgtcagatga ataagaccat cattggtgtg
     2821 tcggtcctca gtgtgcttgt agtatctgtt gtagcagttc tggtctataa gttctatttt
     2881 cacctgatgc ttcttgctgg ctgcataaag tatggtagag gtgaaaacat ctatgatgcc
     2941 tttgttatct actcaagcca ggatgaggac tgggtaagga atgagctagt aaagaattta
     3001 gaagaagggg tgcctccatt tcagctctgc cttcactaca gagactttat tcccggtgtg
     3061 gccattgctg ccaacatcat ccatgaaggt ttccataaaa gccgaaaggt gattgttgtg
     3121 gtgtcccagc acttcatcca gagccgctgg tgtatctttg aatatgagat tgctcagacc
     3181 tggcagtttc tgagcagtcg tgctggtatc atcttcattg tcctgcagaa ggtggagaag
     3241 accctgctca ggcagcaggt ggagctgtac cgccttctca gcaggaacac ttacctggag
     3301 tgggaggaca gtgtcctggg gcggcacatc ttctggagac gactcagaaa agccctgctg
     3361 gatggtaaat catggaatcc agaaggaaca gtgggtacag gatgcaattg gcaggaagca
     3421 acatctatcc tcgagatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3481 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3541 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3601 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgaagtgctt cgcccgctac
     3661 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3721 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3781 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3841 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     3901 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     3961 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4021 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     4081 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     4141 gagctgtaca agtaatctag agggccctat tctatagtgt cacctaaatg ctagagctcg
     4201 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4261 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4321 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4381 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     4441 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4501 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4561 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4621 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4681 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4741 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4801 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4861 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4921 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4981 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5041 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5101 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5161 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     5221 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5281 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5341 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5401 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5461 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5521 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5581 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5641 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5701 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5761 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5821 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5881 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5941 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     6001 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6061 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6121 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     6181 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     6241 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     6301 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6361 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6421 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6481 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6541 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6601 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6661 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6721 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6781 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6841 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6901 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6961 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7021 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7081 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7141 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7201 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7261 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     7321 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7381 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7441 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7501 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7561 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7621 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7681 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7741 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7801 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7861 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7921 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7981 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8041 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8101 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8161 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8221 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8281 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8341 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8401 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8461 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8521 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8581 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
