LOCUS       sequence-417624-        8157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human TRIF protein expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8157)
  TITLE     TREAT-AD Plasmids 2023
REFERENCE   2  (bases 1 to 8157)
  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: pDEST26-hTICAM1-Nluc
FEATURES             Location/Qualifiers
     source          1..8157
                     /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"
     protein_bind    677..701
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             895..906
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2847..2871)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2882..2908
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2939..3454
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(3494..3513)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3538..3586
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3788..4216
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3875..3894)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4085..4106
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4225..4245)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4230..4559
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4410..4545
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4472..4491
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4626..5420
                     /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(4680..4699)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5290..5309
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5596..5717
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5633..5652)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5687..5706
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5766..5782)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5766..5782)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5779..5801)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5790..5806
                     /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(5814..5820,5821..5838,5839..5844))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5859..5880
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5997..6014)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6168..6756)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6248..6267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6927..7718,7719..7787))
                     /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     7550..7569
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7788..7892)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7967..7986)
                     /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 gctagttaag ctagcaacaa gtttgtacaa aaaagcaggc tccaccatgg cctgcacagg
      721 cccatcactt cctagcgcct tcgacattct aggtgcagca ggccaggaca agctcttgta
      781 tctgaagcac aaactgaaga ccccacgccc aggctgccag gggcaggacc tcctgcatgc
      841 catggttctc ctgaagctgg gccaggaaac tgaggccagg atctctctag aggcattgaa
      901 ggccgatgcg gtggcccggc tggtggcccg ccagtgggct ggcgtggaca gcaccgagga
      961 cccagaggag cccccagatg tgtcctgggc tgtggcccgc ttgtaccacc tgctggctga
     1021 ggagaagctg tgccccgcct cgctgcggga cgtggcctac caggaagccg tccgcaccct
     1081 cagctccagg gacgaccacc ggctggggga acttcaggat gaggcccgaa accggtgtgg
     1141 gtgggacatt gctggggatc cagggagcat ccggacgctc cagtccaatc tgggctgcct
     1201 cccaccatcc tcggctttgc cctctgggac caggagcctc ccacgcccca ttgacggtgt
     1261 ttcggactgg agccaagggt gctccctgcg atccactggc agccctgcct ccctggccag
     1321 caacttggaa atcagccagt cccctaccat gcccttcctc agcctgcacc gcagcccaca
     1381 tgggcccagc aagctctgtg acgaccccca ggccagcttg gtgcccgagc ctgtccccgg
     1441 tggctgccag gagcctgagg agatgagctg gccgccatcg ggggagattg ccagcccacc
     1501 agagctgcca agcagcccac ctcctgggct tcccgaagtg gccccagatg caacctccac
     1561 tggcctccct gatacccccg cagctccaga aaccagcacc aactacccag tggagtgcac
     1621 cgaggggtct gcaggccccc agtctctccc cttgcctatt ctggagccgg tcaaaaaccc
     1681 ctgctctgtc aaagaccaga cgccactcca actttctgta gaagatacca cctctccaaa
     1741 taccaagccg tgcccaccta ctcccaccac cccagaaaca tcccctcctc ctcctcctcc
     1801 tcctccttca tctactcctt gttcagctca cctgaccccc tcctccctgt tcccttcctc
     1861 cctggaatca tcatcggaac agaaattcta taactttgtg atcctccacg ccagggcaga
     1921 cgaacacatc gccctgcggg ttcgggagaa gctggaggcc cttggcgtgc ccgacggggc
     1981 caccttctgc gaggatttcc aggtgccggg gcgcggggag ctgagctgcc tgcaggacgc
     2041 catagaccac tcagctttca tcatcctact tctcacctcc aacttcgact gtcgcctgag
     2101 cctgcaccag gtgaaccaag ccatgatgag caacctcacg cgacaggggt cgccagactg
     2161 tgtcatcccc ttcctgcccc tggagagctc cccggcccag ctcagctccg acacggccag
     2221 cctgctctcc gggctggtgc ggctggacga acactcccag atcttcgcca ggaaggtggc
     2281 caacaccttc aagccccaca ggcttcaggc ccgaaaggcc atgtggagga aggaacagga
     2341 cacccgagcc ctgcgggaac agagccaaca cctggacggt gagcggatgc aggcggcggc
     2401 actgaacgca gcctactcag cctacctcca gagctacttg tcctaccagg cacagatgga
     2461 gcagctccag gtggcttttg ggagccacat gtcatttggg actggggcgc cctatggggc
     2521 tcgaatgccc tttgggggcc aggtgcccct gggagccccg ccaccctttc ccacttggcc
     2581 ggggtgcccg cagccgccac ccctgcacgc atggcaggct ggcacccccc caccgccctc
     2641 cccacagcca gcagcctttc cacagtcact gcccttcccg cagtccccag ccttccctac
     2701 ggcctcaccc gcaccccctc agagcccagg gctgcaaccc ctcattatcc accacgcaca
     2761 gatggtacag ctggggctga acaaccacat gtggaaccag agagggtccc aggcgcccga
     2821 ggacaagacg caggaggcag aattggaccc agctttcttg tacaaagtgg ttgatctcga
     2881 gtacccatac gatgttccag attacgctgg agggggtgga tctggagggg gtggatctat
     2941 ggtcttcaca ctcgaagatt tcgttgggga ctggcgacag acagccggct acaacctgga
     3001 ccaagtcctt gaacagggag gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac
     3061 tccgatccaa aggattgtcc tgagcggtga aaatgggctg aagatcgaca tccatgtcat
     3121 catcccgtat gaaggtctga gcggcgacca aatgggccag atcgaaaaaa tttttaaggt
     3181 ggtgtaccct gtggatgatc atcactttaa ggtgatcctg cactatggca cactggtaat
     3241 cgacggggtt acgccgaaca tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt
     3301 gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca aaattatcga
     3361 cgagcgcctg atcaaccccg acggctccct gctgttccga gtaaccatca acggagtgac
     3421 cggctggcgg ctgtgcgaac gcattctggc gtgaggtacc ggttagtaat gagtttaaac
     3481 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     3541 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     3601 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     3661 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     3721 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     3781 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3841 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3901 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3961 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4021 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4081 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4141 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4201 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4261 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4321 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4381 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4441 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4501 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4561 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4621 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4681 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4741 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4801 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4861 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4921 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4981 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5041 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5101 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5161 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5221 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5281 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5341 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5401 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     5461 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5521 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5581 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5641 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5701 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5761 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5821 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5881 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5941 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6001 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6061 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6121 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6181 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6241 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6301 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6361 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6421 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6481 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6541 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6601 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6661 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6721 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6781 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6841 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6901 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6961 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7021 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7081 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7141 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7201 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7261 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7321 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7381 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7441 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7501 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7561 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7621 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7681 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7741 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7801 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7861 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7921 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     7981 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     8041 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     8101 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
