LOCUS       sequence-485462-        7924 bp    DNA     circular SYN 08-SEP-2025
DEFINITION  Expresses VenusFlag-hTRAF3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7924)
  TITLE     TREAT-AD Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7924)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pSCM167-VenusFlag-hTRAF3
FEATURES             Location/Qualifiers
     source          1..7924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    85..309
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     298..315
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        335..353
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     335..352
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    372..396
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2107..2131)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2153..2176)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(join(2216..2926,2927..2929,2930..2932))
                     /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(2258..2279)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2607..2626
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2866..2887
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2948..2967)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2949..2967)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3012..3215)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3041..3061)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3216..3595
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3767..3786
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3861..3965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3966..4034,4035..4826)
                     /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     complement(4184..4203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4997..5585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5486..5505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5739..5756
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5873..5894
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5909..5914,5915..5932,5933..5939)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5947..5963
                     /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)."
     primer_bind     5952..5974
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5971..5987
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5971..5987
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6036..6157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6047..6066)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6101..6120
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6331..7125)
                     /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(6442..6461)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7052..7071
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(7192..7521)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7206..7341
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7260..7279)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7506..7526
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(join(7535..7924,1..39))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7645..7666)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7857..7876
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtg gggatacccc ctagagcccc
       61 agctggttct ttccgcctca gaagccatag agcccaccgc atccccagca tgcctgctat
      121 tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac
      181 acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg
      241 caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag
      301 aaggcacagt cgaggctgat cagcgagctc tagcatttag gtgacactat agaatagggc
      361 cctctagatc aaccactttg tacaagaaag ctgggttcta gggatcgggc agatccgaag
      421 tatccactat gactttaata aaaattgtat catctttaat atatgtccca ttttctagaa
      481 cagtttgggc cacaaagact gggcagccag aggcgatatt catctctcca gtgggcttct
      541 tgaagctgct gctgttgggg tcgggcttga atgcatctcc caaatgacgt cgagaggacc
      601 cctgatccat cagcatgagt gtcactttct gcttaaacgg ccaaggaagc agggcatcat
      661 attctccacg catgatgaca aaaaacagcg acaagtgcgt ccccttcccc atcccgtccc
      721 cgttcaggta gaccctggca cacatcttat agccaaagta accagtgtag aaaggctggc
      781 tgtaaaggga cagggtcttc cccatgacgg cctcctgctt ccgccgcttg tagtcgcgaa
      841 tcttccagat gagcactcca ttgtagctgg cggtctccag gacctggaag cgcaggtcca
      901 tgtcggctag gcggatgtcg tgcacactca gcatctggtc atgccggctc agctgggact
      961 ccagcaggcc tgtgttccga gccacttgcc ccgcgctctt gtccacgctc tccagctcgg
     1021 tcacgcggtt ctggagggac tccacgctgc tcttcatgct gtctgcttcc tcccagttct
     1081 gccggaaggg ccggatctcc ttgtcaagct ccttcagttt ctctgcttgg ctgtctatca
     1141 ctcgctgtaa atgaaggatt ttggattcat tatttcgaag catttccttt tgtctctcaa
     1201 tttcaatttc aaagctacat atctgattgt gcaaactttg tatgctcttg tttttttcta
     1261 cactttcatt ctgcaacaag gaaaccttct tttcgagcga gttgctccac tccttcagca
     1321 ggttgacgtg ctgcacggcg gagctggcct cgtgggcctt gatctgctgg tttgtcccct
     1381 gaaaaacgca gccatagcgc ttaaaactac aggtgctggg ggcattgaca cactctgaca
     1441 agtgtgcact caactcgctc ctcaggagag tctggacgct gcacttgtga gggcaggaca
     1501 ccaccacgca gggacagtcg gtgtcttcgt gtttctgcag cgcgatcatc ggaacctgac
     1561 tcttgcagtg gctgcatgtg gcttcccggt atttacacgc cttctccacg tggtctcgca
     1621 ggtctttcct caagaccttt tctttgcagt caggacgcac acatggaagt tcttcaaaat
     1681 ggcaatcatt ttttaaatgc accagcagat gtcccagcat taactgctct gcacaacctc
     1741 tgctttcatt ccgacaatag atctgaagag ccagaatttc tctcttgcag caattatcct
     1801 taaacacctt atctttaacg atgctctctt gacacgctgt acattttgga cttgaagagc
     1861 tcagcagggc cgccatgcag ctctcgcaga agcggtgccc acactcggtc tgcttcgggc
     1921 tgcacagcac caggtggcac ttctcacact tgtacttgtc ctccacggtc ttcacaaact
     1981 tttccttgta acctccttgt tcagggacaa aaactggcgt cccagcactg cggtcagtgt
     2041 gcagctttag cggcgggtta gtctgcagcg cgccaggaga gtccatcttt ttactcgact
     2101 ccatggtgcc tgcttttttg tacaaacttg ttgataattc ggatccacca cccttgtcat
     2161 cgtcgtcctt gtagtccata agcttagatc caccccctcc agatccaccc cctcccttgt
     2221 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     2281 gatcgcgctt ctcgttgggg tctttgctca gggcggactg gtagctcagg tagtggttgt
     2341 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     2401 gcacgccgcc gtcctcgatg ttgtggcgga tcttgaagtt ggccttgatg ccgttcttct
     2461 gcttgtcggc ggtgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     2521 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     2581 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     2641 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     2701 ggtcggggta gcgggcgaag cactgcaggc cgtagcccag ggtggtcacg agggtgggcc
     2761 agggcacggg cagcttgccg gtggtgcaga tcagcttcag ggtcagcttg ccgtaggtgg
     2821 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     2881 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtaagct
     2941 tgggtctccc tatagtgagt cgtattaatt tcgataagcc agtaagcagt gggttctcta
     3001 gttagccaga gagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
     3061 gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
     3121 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
     3181 ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg
     3241 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
     3301 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
     3361 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     3421 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     3481 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     3541 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgtcaacgc
     3601 gtatatctgg cccgtacatc gcgaagcagc gcaaaacgcc taaccctaag cagattcttc
     3661 atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc gcactactca
     3721 gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc atcagagcag
     3781 attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg acgtcaggtg
     3841 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3901 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3961 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4021 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4081 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4141 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4201 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4261 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4321 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4381 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4441 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4501 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4561 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4621 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4681 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4741 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4801 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4861 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4921 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4981 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5041 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5101 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5161 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5221 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5281 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5341 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5401 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5461 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5521 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5581 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5641 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5701 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5761 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5821 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     5881 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     5941 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     6001 agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca
     6061 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     6121 tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca
     6181 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     6241 acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt
     6301 ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa
     6361 ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca
     6421 ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc
     6481 cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat
     6541 attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc
     6601 cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc
     6661 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
     6721 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
     6781 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
     6841 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
     6901 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc
     6961 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
     7021 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
     7081 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
     7141 tcctgtctct tgatcagatc cgaaaatgga tatacaagct cccgggagct ttttgcaaaa
     7201 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg cagaggcggc
     7261 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     7321 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     7381 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     7441 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     7501 tccacaccct aactgacaca cattccacag aattaattcg cgttaaattt ttgttaaatc
     7561 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     7621 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     7681 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     7741 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     7801 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     7861 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     7921 acca
//
