LOCUS       sequence-392629-        8358 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 8358)
  TITLE     TREAT-AD Plasmids 2023
REFERENCE   2  (bases 1 to 8358)
  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-hTICAM1-VenusFlag
FEATURES             Location/Qualifiers
     source          1..8358
                     /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"
     protein_bind    911..935
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1129..1140
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(3081..3105)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3116..3139
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(3170..3172,3173..3175,3176..3889)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3215..3236)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3476..3495)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3823..3844
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3929..3948)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3973..4021
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4223..4651
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4310..4329)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4520..4541
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4660..4680)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4665..4994
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4845..4980
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4907..4926
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5061..5855
                     /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(5115..5134)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5725..5744
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6031..6152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6068..6087)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6122..6141
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6201..6217)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6201..6217)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6214..6236)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6225..6241
                     /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(6249..6255,6256..6273,6274..6279))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6294..6315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6432..6449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6603..7191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6683..6702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7362..8153,8154..8222))
                     /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     7985..8004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8223..8327)
                     /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 gcttgctagt
      901 taagctagca acaagtttgt acaaaaaagc aggctccacc atggcctgca caggcccatc
      961 acttcctagc gccttcgaca ttctaggtgc agcaggccag gacaagctct tgtatctgaa
     1021 gcacaaactg aagaccccac gcccaggctg ccaggggcag gacctcctgc atgccatggt
     1081 tctcctgaag ctgggccagg aaactgaggc caggatctct ctagaggcat tgaaggccga
     1141 tgcggtggcc cggctggtgg cccgccagtg ggctggcgtg gacagcaccg aggacccaga
     1201 ggagccccca gatgtgtcct gggctgtggc ccgcttgtac cacctgctgg ctgaggagaa
     1261 gctgtgcccc gcctcgctgc gggacgtggc ctaccaggaa gccgtccgca ccctcagctc
     1321 cagggacgac caccggctgg gggaacttca ggatgaggcc cgaaaccggt gtgggtggga
     1381 cattgctggg gatccaggga gcatccggac gctccagtcc aatctgggct gcctcccacc
     1441 atcctcggct ttgccctctg ggaccaggag cctcccacgc cccattgacg gtgtttcgga
     1501 ctggagccaa gggtgctccc tgcgatccac tggcagccct gcctccctgg ccagcaactt
     1561 ggaaatcagc cagtccccta ccatgccctt cctcagcctg caccgcagcc cacatgggcc
     1621 cagcaagctc tgtgacgacc cccaggccag cttggtgccc gagcctgtcc ccggtggctg
     1681 ccaggagcct gaggagatga gctggccgcc atcgggggag attgccagcc caccagagct
     1741 gccaagcagc ccacctcctg ggcttcccga agtggcccca gatgcaacct ccactggcct
     1801 ccctgatacc cccgcagctc cagaaaccag caccaactac ccagtggagt gcaccgaggg
     1861 gtctgcaggc ccccagtctc tccccttgcc tattctggag ccggtcaaaa acccctgctc
     1921 tgtcaaagac cagacgccac tccaactttc tgtagaagat accacctctc caaataccaa
     1981 gccgtgccca cctactccca ccaccccaga aacatcccct cctcctcctc ctcctcctcc
     2041 ttcatctact ccttgttcag ctcacctgac cccctcctcc ctgttccctt cctccctgga
     2101 atcatcatcg gaacagaaat tctataactt tgtgatcctc cacgccaggg cagacgaaca
     2161 catcgccctg cgggttcggg agaagctgga ggcccttggc gtgcccgacg gggccacctt
     2221 ctgcgaggat ttccaggtgc cggggcgcgg ggagctgagc tgcctgcagg acgccataga
     2281 ccactcagct ttcatcatcc tacttctcac ctccaacttc gactgtcgcc tgagcctgca
     2341 ccaggtgaac caagccatga tgagcaacct cacgcgacag gggtcgccag actgtgtcat
     2401 ccccttcctg cccctggaga gctccccggc ccagctcagc tccgacacgg ccagcctgct
     2461 ctccgggctg gtgcggctgg acgaacactc ccagatcttc gccaggaagg tggccaacac
     2521 cttcaagccc cacaggcttc aggcccgaaa ggccatgtgg aggaaggaac aggacacccg
     2581 agccctgcgg gaacagagcc aacacctgga cggtgagcgg atgcaggcgg cggcactgaa
     2641 cgcagcctac tcagcctacc tccagagcta cttgtcctac caggcacaga tggagcagct
     2701 ccaggtggct tttgggagcc acatgtcatt tgggactggg gcgccctatg gggctcgaat
     2761 gccctttggg ggccaggtgc ccctgggagc cccgccaccc tttcccactt ggccggggtg
     2821 cccgcagccg ccacccctgc acgcatggca ggctggcacc cccccaccgc cctccccaca
     2881 gccagcagcc tttccacagt cactgccctt cccgcagtcc ccagccttcc ctacggcctc
     2941 acccgcaccc cctcagagcc cagggctgca acccctcatt atccaccacg cacagatggt
     3001 acagctgggg ctgaacaacc acatgtggaa ccagagaggg tcccaggcgc ccgaggacaa
     3061 gacgcaggag gcagaattgg acccagcttt cttgtacaaa gtggttgatc tcgaggacta
     3121 caaggacgac gatgacaagg gagggggtgg atctggaggg ggtggatcta tggtgagcaa
     3181 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     3241 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     3301 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     3361 cctgggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     3421 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     3481 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3541 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3601 caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     3661 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     3721 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     3781 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3841 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag gtaccggtta
     3901 gtaatgagtt taaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3961 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     4021 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4081 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4141 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     4201 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4261 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4321 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4381 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4441 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4501 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4561 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4621 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4681 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4741 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4801 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4861 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4921 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4981 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     5041 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5101 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5161 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5221 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5281 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5341 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5401 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5461 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5521 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5581 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5641 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5701 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5761 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5821 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     5881 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5941 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     6001 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     6061 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     6121 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     6181 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     6241 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6301 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6361 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6421 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6481 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6541 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6601 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6661 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6721 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6781 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6841 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6901 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6961 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7021 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7081 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7141 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7201 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7261 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7321 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7381 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7441 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7501 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7561 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7621 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7681 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7741 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7801 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7861 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7921 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7981 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     8041 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8101 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8161 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8221 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8281 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8341 aaagtgccac ctgacgtc
//
