LOCUS       sequence-7719-ex        8796 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 8796)
  TITLE     TLR Signaling Plasmids
REFERENCE   2  (bases 1 to 8796)
  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-TLR3-CFP
FEATURES             Location/Qualifiers
     source          1..8796
                     /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(3613..3615,3616..3618,3619..4332)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3658..3679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3919..3938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4266..4287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4348..4366)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4349..4366)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4386..4403)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4392..4616
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4662..5090
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4749..4768)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4959..4980
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5099..5119)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5104..5434
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5285..5420
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5347..5366
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5501..6295
                     /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(5555..5574)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6165..6184
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6469..6590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6506..6525)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6560..6579
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6639..6655)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6639..6655)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6652..6674)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6663..6679
                     /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(6687..6693,6694..6711,6712..6717))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6732..6753
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6870..6887)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7041..7629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7121..7140)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7800..8591,8592..8660))
                     /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     8423..8442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8661..8765)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttatgaga
      901 cagactttgc cttgtatcta cttttggggg ggcctcttgc cctttgggat gctgtgtgca
      961 tcctccacca ccaagtgcac tgttagccat gaagttgctg actgcagcca cctgaagttg
     1021 actcaggtac ccgatgatct acccacaaac ataacagtgt tgaaccttac ccataatcaa
     1081 ctcagaagat taccagccgc caacttcaca aggtatagcc agctaactag cttggatgta
     1141 ggatttaaca ccatctcaaa actggagcca gaattgtgcc agaaacttcc catgttaaaa
     1201 gttttgaacc tccagcacaa tgagctatct caactttctg ataaaacctt tgccttctgc
     1261 acgaatttga ctgaactcca tctcatgtcc aactcaatcc agaaaattaa aaataatccc
     1321 tttgtcaagc agaagaattt aatcacatta gatctgtctc ataatggctt gtcatctaca
     1381 aaattaggaa ctcaggttca gctggaaaat ctccaagagc ttctattatc aaacaataaa
     1441 attcaagcgc taaaaagtga agaactggat atctttgcca attcatcttt aaaaaaatta
     1501 gagttgtcat cgaatcaaat taaagagttt tctccagggt gttttcacgc aattggaaga
     1561 ttatttggcc tctttctgaa caatgtccag ctgggtccca gccttacaga gaagctatgt
     1621 ttggaattag caaacacaag cattcggaat ctgtctctga gtaacagcca gctgtccacc
     1681 accagcaata caactttctt gggactaaag tggacaaatc tcactatgct cgatctttcc
     1741 tacaacaact taaatgtggt tggtaacgat tcctttgctt ggcttccaca actagaatat
     1801 ttcttcctag agtataataa tatacagcat ttgttttctc actctttgca cgggcttttc
     1861 aatgtgaggt acctgaattt gaaacggtct tttactaaac aaagtatttc ccttgcctca
     1921 ctccccaaga ttgatgattt ttcttttcag tggctaaaat gtttggagca ccttaacatg
     1981 gaagataatg atattccagg cataaaaagc aatatgttca caggattgat aaacctgaaa
     2041 tacttaagtc tatccaactc ctttacaagt ttgcgaactt tgacaaatga aacatttgta
     2101 tcacttgctc attctccctt acacatactc aacctaacca agaataaaat ctcaaaaata
     2161 gagagtgatg ctttctcttg gttgggccac ctagaagtac ttgacctggg ccttaatgaa
     2221 attgggcaag aactcacagg ccaggaatgg agaggtctag aaaatatttt cgaaatctat
     2281 ctttcctaca acaagtacct gcagctgact aggaactcct ttgccttggt cccaagcctt
     2341 caacgactga tgctccgaag ggtggccctt aaaaatgtgg atagctctcc ttcaccattc
     2401 cagcctcttc gtaacttgac cattctggat ctaagcaaca acaacatagc caacataaat
     2461 gatgacatgt tggagggtct tgagaaacta gaaattctcg atttgcagca taacaactta
     2521 gcacggctct ggaaacacgc aaaccctggt ggtcccattt atttcctaaa gggtctgtct
     2581 cacctccaca tccttaactt ggagtccaac ggctttgacg agatcccagt tgaggtcttc
     2641 aaggatttat ttgaactaaa gatcatcgat ttaggattga ataatttaaa cacacttcca
     2701 gcatctgtct ttaataatca ggtgtctcta aagtcattga accttcagaa gaatctcata
     2761 acatccgttg agaagaaggt tttcgggcca gctttcagga acctgactga gttagatatg
     2821 cgctttaatc cctttgattg cacgtgtgaa agtattgcct ggtttgttaa ttggattaac
     2881 gagacccata ccaacatccc tgagctgtca agccactacc tttgcaacac tccacctcac
     2941 tatcatgggt tcccagtgag actttttgat acatcatctt gcaaagacag tgcccccttt
     3001 gaactctttt tcatgatcaa taccagtatc ctgttgattt ttatctttat tgtacttctc
     3061 atccactttg agggctggag gatatctttt tattggaatg tttcagtaca tcgagttctt
     3121 ggtttcaaag aaatagacag acagacagaa cagtttgaat atgcagcata tataattcat
     3181 gcctataaag ataaggattg ggtctgggaa catttctctt caatggaaaa ggaagaccaa
     3241 tctctcaaat tttgtctgga agaaagggac tttgaggcgg gtgtttttga actagaagca
     3301 attgttaaca gcatcaaaag aagcagaaaa attatttttg ttataacaca ccatctatta
     3361 aaagacccat tatgcaaaag attcaaggta catcatgcag ttcaacaagc tattgaacaa
     3421 aatctggatt ccattatatt ggttttcctt gaggagattc cagattataa actgaaccat
     3481 gcactctgtt tgcgaagagg aatgtttaaa tctcactgca tcttgaactg gccagttcag
     3541 aaagaacgga taggtgcctt tcgtcataaa ttgcaagtag cacttggatc caaaaactct
     3601 gtacatctcg agatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3661 gtcgagctgg acggcgacgt aaacggccac aggttcagcg tgtccggcga gggcgagggc
     3721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     3781 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag ccgctacccc
     3841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3901 cgtaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     4021 atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat caccgccgac
     4081 aagcagaaga acggcatcaa ggcccacttc aagatccgcc acaacatcga ggacggcagc
     4141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     4201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     4261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     4321 ctgtacaagt aatctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg
     4381 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     4441 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     4501 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     4561 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     4621 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     4681 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4741 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4801 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     4861 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4921 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4981 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     5041 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     5101 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca
     5161 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     5221 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     5281 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     5341 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     5401 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     5461 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     5521 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     5581 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     5641 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     5701 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5761 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5821 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5881 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5941 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     6001 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     6061 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     6121 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     6181 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     6241 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     6301 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     6361 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     6421 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     6481 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6541 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     6601 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     6661 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6721 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6781 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6841 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6901 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6961 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     7021 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     7081 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     7141 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     7201 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     7261 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7321 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7381 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7441 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     7501 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7561 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7621 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7681 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7741 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7801 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7861 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7921 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7981 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8041 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8101 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8161 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8221 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     8281 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8341 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8401 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8461 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8521 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8581 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8641 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8701 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8761 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
