LOCUS       sequence-375252-        7491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of TDP-43 YFP in mammalian cells.
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 YFP
FEATURES             Location/Qualifiers
     source          1..7491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(20..39)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        211..590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        591..794
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     745..765
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     839..858
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        839..857
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    887..911
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2183..2893
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(2222..2243)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2483..2502)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2830..2851
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2907..2931)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2957..2998
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3038..3057)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3082..3130
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3332..3760
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3419..3438)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3629..3650
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3769..3789)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3774..4103
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3954..4089
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4016..4035
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4170..4964
                     /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(4224..4243)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4834..4853
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5140..5261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5177..5196)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5231..5250
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5310..5326)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5310..5326)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5323..5345)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5334..5350
                     /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(5358..5364,5365..5382,5383..5388))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5403..5424
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5541..5558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5712..6300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5792..5811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6471..7262,7263..7331))
                     /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     7094..7113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7332..7436)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
       61 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
      121 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
      181 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
      241 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      301 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      361 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      421 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      481 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      541 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      601 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      661 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      721 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      781 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
      841 atacgactca ctatagggag acccaagctg gctagttaag ctatcaacaa gtttgtacaa
      901 aaaagcaggc ttcgaaggag ataacaaaat gtctgaatat attcgggtaa ccgaagatga
      961 gaacgatgag cccattgaaa taccatcgga agacgatggg acggtgctgc tctccacggt
     1021 tacagcccag tttccagggg cgtgtgggct tcgctacagg aatccagtgt ctcagtgtat
     1081 gagaggtgtc cggctggtag aaggaattct gcatgcccca gatgctggct ggggaaatct
     1141 ggtgtatgtt gtcaactatc caaaagataa caaaagaaaa atggatgaga cagatgcttc
     1201 atcagcagtg aaagtgaaaa gagcagtcca gaaaacatcc gatttaatag tgttgggtct
     1261 cccatggaaa acaaccgaac aggacctgaa agagtatttt agtacctttg gagaagttct
     1321 tatggtgcag gtcaagaaag atcttaagac tggtcattca aaggggtttg gctttgttcg
     1381 ttttacggaa tatgaaacac aagtgaaagt aatgtcacag cgacatatga tagatggacg
     1441 atggtgtgac tgcaaacttc ctaattctaa gcaaagccaa gatgagcctt tgagaagcag
     1501 aaaagtgttt gtggggcgct gtacagagga catgactgag gatgagctgc gggagttctt
     1561 ctctcagtac ggggatgtga tggatgtctt catccccaag ccattcaggg cctttgcctt
     1621 tgttacattt gcagatgatc agattgcgca gtctctttgt ggagaggact tgatcattaa
     1681 aggaatcagc gttcatatat ccaatgccga acctaagcac aatagcaata gacagttaga
     1741 aagaagtgga agatttggtg gtaatccagg tggctttggg aatcagggtg gatttggtaa
     1801 tagcagaggg ggtggagctg gtttgggaaa caatcaaggt agtaatatgg gtggtgggat
     1861 gaactttggt gcgttcagca ttaatccagc catgatggct gccgcccagg cagcactaca
     1921 gagcagttgg ggtatgatgg gcatgttagc cagccagcag aaccagtcag gcccatcggg
     1981 taataaccaa aaccaaggca acatgcagag ggagccaaac caggccttcg gttctggaaa
     2041 taactcttat agtggctcta attctggtgc agcaattggt tggggatcag catccaatgc
     2101 agggtcgggc agtggtttta atggaggctt tggctcaagc atggattcta agtcttctgg
     2161 ctggggaatg aagcttatcg atagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     2221 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2281 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     2341 cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgcagtgct tcgcccgcta
     2401 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2461 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2521 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2581 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     2641 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     2701 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2761 gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa
     2821 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2881 cgagctgtac aagagatcta tctaggaccc agctttcttg tacaaagtgg ttgatctaga
     2941 gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     3001 taccggttag taatgagttt aaacggggga ggctaactga aacacggaag gagacaatac
     3061 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     3121 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     3181 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     3241 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     3301 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3361 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3421 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3481 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3541 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3601 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3661 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3721 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3781 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3841 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3901 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3961 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4021 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4081 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4141 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4201 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4261 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4321 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4381 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4441 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4501 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4561 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4621 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4681 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4741 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4801 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4861 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4921 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4981 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5041 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5101 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5161 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5221 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5281 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5341 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5401 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5461 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5521 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5581 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5641 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5701 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5761 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5821 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5881 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5941 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6001 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6061 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6121 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6181 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6241 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6301 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6361 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6421 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6481 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6541 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6601 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6661 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6721 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6781 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6841 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6901 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6961 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7021 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7081 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7141 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7201 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7261 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7321 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7381 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7441 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat c
//
