LOCUS       sequence-375251-        7491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expresion of  TDP-43 NLS1 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 NLS1 YFP
FEATURES             Location/Qualifiers
     source          1..7491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..137
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     64..83
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             218..1012
                     /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(272..291)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     882..901
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1188..1309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1225..1244)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1279..1298
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1358..1374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1358..1374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1371..1393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1382..1398
                     /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(1406..1412,1413..1430,1431..1436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1451..1472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1589..1606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1760..2348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1840..1859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2519..3310,3311..3379))
                     /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     3142..3161
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3380..3484)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3559..3578)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3750..4129
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4130..4333
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4284..4304
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4378..4397
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4378..4396
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4426..4450
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5722..6432
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(5761..5782)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6022..6041)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6369..6390
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(6446..6470)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6496..6537
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(6577..6596)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6621..6669
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6871..7299
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6958..6977)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7168..7189
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7308..7328)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(7313..7491,1..151)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
       61 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
      121 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
      181 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
      241 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
      301 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
      361 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
      421 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
      481 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
      541 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
      601 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
      661 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
      721 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
      781 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
      841 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
      901 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
      961 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     1021 ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     1081 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     1141 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     1201 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1261 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     1321 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1441 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1501 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1561 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1621 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1681 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1741 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1801 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1861 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1921 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1981 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2041 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2101 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2161 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2221 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2281 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2341 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2401 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2461 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2521 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2581 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2641 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2701 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2761 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2821 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2881 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     2941 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3001 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3061 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3121 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3181 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3241 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3301 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3361 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3421 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3481 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     3541 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     3601 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     3661 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     3721 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
     3781 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     3841 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     3901 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     3961 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     4021 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     4081 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     4141 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     4201 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     4261 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     4321 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
     4381 tacgactcac tatagggaga cccaagctgg ctagttaagc tatcaacaag tttgtacaaa
     4441 aaagcaggct tcgaaggaga taacaaaatg tctgaatata ttcgggtaac cgaagatgag
     4501 aacgatgagc ccattgaaat accatcggaa gacgatggga cggtgctgct ctccacggtt
     4561 acagcccagt ttccaggggc gtgtgggctt cgctacagga atccagtgtc tcagtgtatg
     4621 agaggtgtcc ggctggtaga aggaattctg catgccccag atgctggctg gggaaatctg
     4681 gtgtatgttg tcaactatcc aaaagataac gcagcagcaa tggatgagac agatgcttca
     4741 tcagcagtga aagtgaaaag agcagtccag aaaacatccg atttaatagt gttgggtctc
     4801 ccatggaaaa caaccgaaca ggacctgaaa gagtatttta gtacctttgg agaagttctt
     4861 atggtgcagg tcaagaaaga tcttaagact ggtcattcaa aggggtttgg ctttgttcgt
     4921 tttacggaat atgaaacaca agtgaaagta atgtcacagc gacatatgat agatggacga
     4981 tggtgtgact gcaaacttcc taattctaag caaagccaag atgagccttt gagaagcaga
     5041 aaagtgtttg tggggcgctg tacagaggac atgactgagg atgagctgcg ggagttcttc
     5101 tctcagtacg gggatgtgat ggatgtcttc atccccaagc cattcagggc ctttgccttt
     5161 gttacatttg cagatgatca gattgcgcag tctctttgtg gagaggactt gatcattaaa
     5221 ggaatcagcg ttcatatatc caatgccgaa cctaagcaca atagcaatag acagttagaa
     5281 agaagtggaa gatttggtgg taatccaggt ggctttggga atcagggtgg atttggtaat
     5341 agcagagggg gtggagctgg tttgggaaac aatcaaggta gtaatatggg tggtgggatg
     5401 aactttggtg cgttcagcat taatccagcc atgatggctg ccgcccaggc agcactacag
     5461 agcagttggg gtatgatggg catgttagcc agccagcaga accagtcagg cccatcgggt
     5521 aataaccaaa accaaggcaa catgcagagg gagccaaacc aggccttcgg ttctggaaat
     5581 aactcttata gtggctctaa ttctggtgca gcaattggtt ggggatcagc atccaatgca
     5641 gggtcgggca gtggttttaa tggaggcttt ggctcaagca tggattctaa gtcttctggc
     5701 tggggaatga agcttatcga tagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     5761 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     5821 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     5881 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac
     5941 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     6001 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     6061 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     6121 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     6181 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     6241 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     6301 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     6361 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     6421 gagctgtaca agagatctat ctaggaccca gctttcttgt acaaagtggt tgatctagag
     6481 ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgcgt
     6541 accggttagt aatgagttta aacgggggag gctaactgaa acacggaagg agacaatacc
     6601 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt
     6661 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     6721 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     6781 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc
     6841 tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     6901 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     6961 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     7021 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     7081 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     7141 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     7201 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     7261 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     7321 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     7381 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     7441 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc c
//
