LOCUS       sequence-392627-        8359 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 8359)
  TITLE     TREAT-AD Plasmids 2023
REFERENCE   2  (bases 1 to 8359)
  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-VenusFlag-hTICAM1
FEATURES             Location/Qualifiers
     source          1..8359
                     /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(898..900,901..903,904..1614)
                     /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(943..964)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1204..1223)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1551..1572
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1654..1677
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1699..1723
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1917..1928
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(3869..3893)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(3912..3930)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3913..3930)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3950..3967)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3956..4180
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4226..4654
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4313..4332)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4523..4544
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4663..4683)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4668..4997
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4848..4983
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4910..4929
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5064..5858
                     /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(5118..5137)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5728..5747
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6032..6153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6069..6088)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6123..6142
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6202..6218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6202..6218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6215..6237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6226..6242
                     /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(6250..6256,6257..6274,6275..6280))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6295..6316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6433..6450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6604..7192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6684..6703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7363..8154,8155..8223))
                     /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     7986..8005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8224..8328)
                     /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 gcttaccatg
      901 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      961 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1021 aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1081 gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
     1141 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1201 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1261 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1321 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
     1381 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
     1441 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1501 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1561 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggaggg
     1621 ggtggatctg gagggggtgg atctaagctt atggactaca aggacgacga tgacaagggt
     1681 ggtggatccg aattatcaac aagtttgtac aaaaaagcag gctccaccat ggcctgcaca
     1741 ggcccatcac ttcctagcgc cttcgacatt ctaggtgcag caggccagga caagctcttg
     1801 tatctgaagc acaaactgaa gaccccacgc ccaggctgcc aggggcagga cctcctgcat
     1861 gccatggttc tcctgaagct gggccaggaa actgaggcca ggatctctct agaggcattg
     1921 aaggccgatg cggtggcccg gctggtggcc cgccagtggg ctggcgtgga cagcaccgag
     1981 gacccagagg agcccccaga tgtgtcctgg gctgtggccc gcttgtacca cctgctggct
     2041 gaggagaagc tgtgccccgc ctcgctgcgg gacgtggcct accaggaagc cgtccgcacc
     2101 ctcagctcca gggacgacca ccggctgggg gaacttcagg atgaggcccg aaaccggtgt
     2161 gggtgggaca ttgctgggga tccagggagc atccggacgc tccagtccaa tctgggctgc
     2221 ctcccaccat cctcggcttt gccctctggg accaggagcc tcccacgccc cattgacggt
     2281 gtttcggact ggagccaagg gtgctccctg cgatccactg gcagccctgc ctccctggcc
     2341 agcaacttgg aaatcagcca gtcccctacc atgcccttcc tcagcctgca ccgcagccca
     2401 catgggccca gcaagctctg tgacgacccc caggccagct tggtgcccga gcctgtcccc
     2461 ggtggctgcc aggagcctga ggagatgagc tggccgccat cgggggagat tgccagccca
     2521 ccagagctgc caagcagccc acctcctggg cttcccgaag tggccccaga tgcaacctcc
     2581 actggcctcc ctgatacccc cgcagctcca gaaaccagca ccaactaccc agtggagtgc
     2641 accgaggggt ctgcaggccc ccagtctctc cccttgccta ttctggagcc ggtcaaaaac
     2701 ccctgctctg tcaaagacca gacgccactc caactttctg tagaagatac cacctctcca
     2761 aataccaagc cgtgcccacc tactcccacc accccagaaa catcccctcc tcctcctcct
     2821 cctcctcctt catctactcc ttgttcagct cacctgaccc cctcctccct gttcccttcc
     2881 tccctggaat catcatcgga acagaaattc tataactttg tgatcctcca cgccagggca
     2941 gacgaacaca tcgccctgcg ggttcgggag aagctggagg cccttggcgt gcccgacggg
     3001 gccaccttct gcgaggattt ccaggtgccg gggcgcgggg agctgagctg cctgcaggac
     3061 gccatagacc actcagcttt catcatccta cttctcacct ccaacttcga ctgtcgcctg
     3121 agcctgcacc aggtgaacca agccatgatg agcaacctca cgcgacaggg gtcgccagac
     3181 tgtgtcatcc ccttcctgcc cctggagagc tccccggccc agctcagctc cgacacggcc
     3241 agcctgctct ccgggctggt gcggctggac gaacactccc agatcttcgc caggaaggtg
     3301 gccaacacct tcaagcccca caggcttcag gcccgaaagg ccatgtggag gaaggaacag
     3361 gacacccgag ccctgcggga acagagccaa cacctggacg gtgagcggat gcaggcggcg
     3421 gcactgaacg cagcctactc agcctacctc cagagctact tgtcctacca ggcacagatg
     3481 gagcagctcc aggtggcttt tgggagccac atgtcatttg ggactggggc gccctatggg
     3541 gctcgaatgc cctttggggg ccaggtgccc ctgggagccc cgccaccctt tcccacttgg
     3601 ccggggtgcc cgcagccgcc acccctgcac gcatggcagg ctggcacccc cccaccgccc
     3661 tccccacagc cagcagcctt tccacagtca ctgcccttcc cgcagtcccc agccttccct
     3721 acggcctcac ccgcaccccc tcagagccca gggctgcaac ccctcattat ccaccacgca
     3781 cagatggtac agctggggct gaacaaccac atgtggaacc agagagggtc ccaggcgccc
     3841 gaggacaaga cgcaggaggc agaattggac ccagctttct tgtacaaagt ggttgatcta
     3901 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     3961 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4021 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4081 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4141 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     4201 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4261 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4321 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4381 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4441 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4501 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4561 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4621 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4681 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4741 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4801 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4861 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4921 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4981 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     5041 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5101 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5161 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5221 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5281 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5341 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5401 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5461 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5521 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5581 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5641 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5701 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5761 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5821 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5881 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5941 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6001 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6061 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6121 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6181 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6241 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6301 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6361 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6421 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6481 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6541 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6601 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6661 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6721 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6781 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6841 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6901 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6961 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7021 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7081 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7141 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7201 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7261 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7321 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7381 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7441 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7501 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7561 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7621 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7681 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7741 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7801 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7861 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7921 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7981 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8041 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8101 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8161 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8221 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8281 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8341 aaaagtgcca cctgacgtc
//
