LOCUS       sequence-485459-        7577 bp    DNA     circular SYN 08-SEP-2025
DEFINITION  Expresses CNluc-HA-hTRAF3-V5 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7577)
  TITLE     TREAT-AD Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7577)
  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            "Unpublished"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: pcDNA3.2-CNluc-HA-hTRAF3-V5
FEATURES             Location/Qualifiers
     source          1..7577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(17..36)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     71..90
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(150..166)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(150..166)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(163..185)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    174..190
                     /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(198..204,205..222,223..228))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    243..264
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(381..398)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(552..1140)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(632..651)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1311..2102,2103..2171))
                     /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     1934..1953
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2172..2276)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2351..2370)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2542..2921
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2922..3125
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3076..3096
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3170..3189
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3170..3188
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3553..3579
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    3589..3613
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5324..5348)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5374..5415
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(5455..5474)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5499..5547
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5749..6177
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5836..5855)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6046..6067
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6186..6206)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6191..6520
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6371..6506
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6433..6452
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6587..7381
                     /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(6641..6660)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7251..7270
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    join(7557..7577,1..101)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
       61 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
      121 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
      181 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
      241 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
      301 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
      361 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
      421 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      481 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      541 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      601 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      661 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      721 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
      781 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      841 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      901 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      961 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1021 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1081 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1141 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1201 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     1261 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     1321 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1381 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1441 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     1501 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     1561 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     1621 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     1681 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1741 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     1801 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1861 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1921 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1981 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     2041 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     2101 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     2161 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2221 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     2281 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     2341 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     2401 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     2461 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     2521 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     2581 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     2641 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     2701 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     2761 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     2821 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     2881 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     2941 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     3001 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     3061 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     3121 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
     3181 actataggga gacccaagct taccatgctg agcggcgacc aaatgggcca gatcgaaaaa
     3241 atttttaagg tggtgtaccc tgtggatgat catcacttta aggtgatcct gcactatggc
     3301 acactggtaa tcgacggggt tacgccgaac atgatcgact atttcggacg gccgtatgaa
     3361 ggcatcgccg tgttcgacgg caaaaagatc actgtaacag ggaccctgtg gaacggcaac
     3421 aaaattatcg acgagcgcct gatcaacccc gacggctccc tgctgttccg agtaaccatc
     3481 aacggagtga ccggctggcg gctgtgcgaa cgcattctgg cgggaggggg tggatctgga
     3541 gggggtggat cttacccata cgatgttcca gattacgctg cgctagcaac aagtttgtac
     3601 aaaaaagcag gcaccatgga gtcgagtaaa aagatggact ctcctggcgc gctgcagact
     3661 aacccgccgc taaagctgca cactgaccgc agtgctggga cgccagtttt tgtccctgaa
     3721 caaggaggtt acaaggaaaa gtttgtgaag accgtggagg acaagtacaa gtgtgagaag
     3781 tgccacctgg tgctgtgcag cccgaagcag accgagtgtg ggcaccgctt ctgcgagagc
     3841 tgcatggcgg ccctgctgag ctcttcaagt ccaaaatgta cagcgtgtca agagagcatc
     3901 gttaaagata aggtgtttaa ggataattgc tgcaagagag aaattctggc tcttcagatc
     3961 tattgtcgga atgaaagcag aggttgtgca gagcagttaa tgctgggaca tctgctggtg
     4021 catttaaaaa atgattgcca ttttgaagaa cttccatgtg tgcgtcctga ctgcaaagaa
     4081 aaggtcttga ggaaagacct gcgagaccac gtggagaagg cgtgtaaata ccgggaagcc
     4141 acatgcagcc actgcaagag tcaggttccg atgatcgcgc tgcagaaaca cgaagacacc
     4201 gactgtccct gcgtggtggt gtcctgccct cacaagtgca gcgtccagac tctcctgagg
     4261 agcgagttga gtgcacactt gtcagagtgt gtcaatgccc ccagcacctg tagttttaag
     4321 cgctatggct gcgtttttca ggggacaaac cagcagatca aggcccacga ggccagctcc
     4381 gccgtgcagc acgtcaacct gctgaaggag tggagcaact cgctcgaaaa gaaggtttcc
     4441 ttgttgcaga atgaaagtgt agaaaaaaac aagagcatac aaagtttgca caatcagata
     4501 tgtagctttg aaattgaaat tgagagacaa aaggaaatgc ttcgaaataa tgaatccaaa
     4561 atccttcatt tacagcgagt gatagacagc caagcagaga aactgaagga gcttgacaag
     4621 gagatccggc ccttccggca gaactgggag gaagcagaca gcatgaagag cagcgtggag
     4681 tccctccaga accgcgtgac cgagctggag agcgtggaca agagcgcggg gcaagtggct
     4741 cggaacacag gcctgctgga gtcccagctg agccggcatg accagatgct gagtgtgcac
     4801 gacatccgcc tagccgacat ggacctgcgc ttccaggtcc tggagaccgc cagctacaat
     4861 ggagtgctca tctggaagat tcgcgactac aagcggcgga agcaggaggc cgtcatgggg
     4921 aagaccctgt ccctttacag ccagcctttc tacactggtt actttggcta taagatgtgt
     4981 gccagggtct acctgaacgg ggacgggatg gggaagggga cgcacttgtc gctgtttttt
     5041 gtcatcatgc gtggagaata tgatgccctg cttccttggc cgtttaagca gaaagtgaca
     5101 ctcatgctga tggatcaggg gtcctctcga cgtcatttgg gagatgcatt caagcccgac
     5161 cccaacagca gcagcttcaa gaagcccact ggagagatga atatcgcctc tggctgccca
     5221 gtctttgtgg cccaaactgt tctagaaaat gggacatata ttaaagatga tacaattttt
     5281 attaaagtca tagtggatac ttcggatctg cccgatccct agaacccagc tttcttgtac
     5341 aaagtggttg atctcgaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc
     5401 ggtctcgatt ctacgggtac cggttagtaa tgagtttaaa cgggggaggc taactgaaac
     5461 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5521 aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5581 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5641 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     5701 gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta
     5761 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5821 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5881 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5941 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     6001 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6061 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6121 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     6181 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     6241 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     6301 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     6361 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6421 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     6481 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     6541 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     6601 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     6661 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     6721 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     6781 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     6841 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     6901 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     6961 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     7021 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     7081 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     7141 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     7201 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     7261 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     7321 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     7381 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     7441 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     7501 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     7561 tgtttattgc agcttat
//
