LOCUS       sequence-300585-        7491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expresion of TDP-43 5F-L YFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7491)
  AUTHORS   Elden AC, Kim HJ, Hart MP, Chen-Plotkin AS, Johnson BS, Fang X, 
            Armakola M, Geser F, Greene R, Lu MM, Padmanabhan A, Clay-Falcone D,
            McCluskey L, Elman L, Juhr D, Gruber PJ, Rub U, Auburger G, 
            Trojanowski JQ, Lee VM, Van Deerlin VM, Bonini NM, Gitler AD
  TITLE     Ataxin-2 intermediate-length polyglutamine expansions are associated
            with increased risk for ALS.
  JOURNAL   Nature. 2010 Aug 26;466(7310):1069-75. doi: 10.1038/nature09320.
  PUBMED    20740007
REFERENCE   2  (bases 1 to 7491)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2010-08-26- 26"; volume: "466"; issue: "7310"; 
            pages: "1069-75"
COMMENT     Sequence Label: pcDNA3.2 TDP-43 5F-L YFP
FEATURES             Location/Qualifiers
     source          1..7491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        171..550
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        551..754
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     705..725
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     799..818
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        799..817
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    847..871
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2143..2853
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(2182..2203)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2443..2462)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2790..2811
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2867..2891)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2917..2958
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2998..3017)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3042..3090
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3292..3720
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3379..3398)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3589..3610
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3729..3749)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3734..4063
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3914..4049
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3976..3995
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4130..4924
                     /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(4184..4203)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4794..4813
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5100..5221
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5137..5156)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5191..5210
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5270..5286)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5270..5286)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5283..5305)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5294..5310
                     /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(5318..5324,5325..5342,5343..5348))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5363..5384
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5501..5518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5672..6260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5752..5771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6431..7222,7223..7291))
                     /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     7054..7073
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7292..7396)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7471..7490)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
       61 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
      121 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
      181 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      241 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      301 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      361 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      421 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      481 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      541 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      601 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      661 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      721 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc
      781 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg gctagttaag
      841 ctatcaacaa gtttgtacaa aaaagcaggc ttcgaaggag ataacaaaat gtctgaatat
      901 attcgggtaa ccgaagatga gaacgatgag cccattgaaa taccatcgga agacgatggg
      961 acggtgctgc tctccacggt tacagcccag tttccagggg cgtgtgggct tcgctacagg
     1021 aatccagtgt ctcagtgtat gagaggtgtc cggctggtag aaggaattct gcatgcccca
     1081 gatgctggct ggggaaatct ggtgtatgtt gtcaactatc caaaagataa caaaagaaaa
     1141 atggatgaga cagatgcttc atcagcagtg aaagtgaaaa gagcagtcca gaaaacatcc
     1201 gatttaatag tgttgggtct cccatggaaa acaaccgaac aggacctgaa agagtatttt
     1261 agtacctttg gagaagttct tatggtgcag gtcaagaaag atcttaagac tggtcattca
     1321 aaggggttag gcttagttcg ttttacggaa tatgaaacac aagtgaaagt aatgtcacag
     1381 cgacatatga tagatggacg atggtgtgac tgcaaacttc ctaattctaa gcaaagccaa
     1441 gatgagcctt tgagaagcag aaaagtgtta gtggggcgct gtacagagga catgactgag
     1501 gatgagctgc gggagttctt ctctcagtac ggggatgtga tggatgtctt catccccaag
     1561 ccattcaggg ccttagcctt agttacattt gcagatgatc agattgcgca gtctctttgt
     1621 ggagaggact tgatcattaa aggaatcagc gttcatatat ccaatgccga acctaagcac
     1681 aatagcaata gacagttaga aagaagtgga agatttggtg gtaatccagg tggctttggg
     1741 aatcagggtg gatttggtaa tagcagaggg ggtggagctg gtttgggaaa caatcaaggt
     1801 agtaatatgg gtggtgggat gaactttggt gcgttcagca ttaatccagc catgatggct
     1861 gccgcccagg cagcactaca gagcagttgg ggtatgatgg gcatgttagc cagccagcag
     1921 aaccagtcag gcccatcggg taataaccaa aaccaaggca acatgcagag ggagccaaac
     1981 caggccttcg gttctggaaa taactcttat agtggctcta attctggtgc agcaattggt
     2041 tggggatcag catccaatgc agggtcgggc agtggtttta atggaggctt tggctcaagc
     2101 atggattcta agtcttctgg ctggggaatg aagcttatcg atagcaaggg cgaggagctg
     2161 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2221 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2281 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc
     2341 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2401 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2461 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2521 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     2581 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     2641 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     2701 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg
     2761 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2821 gggatcactc tcggcatgga cgagctgtac aagagatcta tctaggaccc agctttcttg
     2881 tacaaagtgg ttgatctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc
     2941 ctcggtctcg attctacgcg taccggttag taatgagttt aaacggggga ggctaactga
     3001 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     3061 taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     3121 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     3181 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     3241 caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct
     3301 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3361 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3421 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3481 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3541 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3601 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3661 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3721 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3781 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3841 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3901 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3961 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4021 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     4081 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4141 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4201 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4261 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4321 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4381 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4441 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4501 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4561 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4621 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4681 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4741 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4801 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4861 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4921 ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4981 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5041 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5101 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5161 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5221 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5281 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5341 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5401 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5461 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5521 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5581 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5641 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5701 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5761 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5821 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5881 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5941 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6001 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6061 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6121 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6181 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6241 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6301 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6361 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6421 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6481 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6541 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6601 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6661 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6721 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6781 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6841 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6901 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6961 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7021 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7081 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7141 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7201 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7261 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7321 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7381 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7441 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg c
//
