LOCUS       sequence-258225-        4539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4539)
  AUTHORS   Johnson BS, McCaffery JM, Lindquist S, Gitler AD
  TITLE     A yeast TDP-43 proteinopathy model: Exploring the molecular 
            determinants of TDP-43 aggregation and cellular toxicity.
  JOURNAL   Proc Natl Acad Sci U S A. 2008 Apr 29. 105(17):6439-44.
  PUBMED    18434538
REFERENCE   2  (bases 1 to 4539)
  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"; journalName: "Proc Natl 
            Acad Sci U S A. 2008 Apr 29. 105(17):6439-44."
COMMENT     Sequence Label: pDONR TDP43 WT YFP
FEATURES             Location/Qualifiers
     source          1..4539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..87)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     136..158
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     150..167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     151..167
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    183..282
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1554..2264
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(1593..1614)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1854..1873)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2201..2222
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2278..2377)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2394..2413)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2395..2413)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2418..2434)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2418..2434)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2536..2555)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2547..3356
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2614..2633)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3503..4091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3992..4011
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4245..4262
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4421..4448)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
       61 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
      121 tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc
      181 cccaaataat gattttattt tgactgatag tgacctgttc gttgcaacaa attgatgagc
      241 aatgcttttt tataatgcca actttgtaca aaaaagcagg cttcgaagga gataacaaaa
      301 tgtctgaata tattcgggta accgaagatg agaacgatga gcccattgaa ataccatcgg
      361 aagacgatgg gacggtgctg ctctccacgg ttacagccca gtttccaggg gcgtgtgggc
      421 ttcgctacag gaatccagtg tctcagtgta tgagaggtgt ccggctggta gaaggaattc
      481 tgcatgcccc agatgctggc tggggaaatc tggtgtatgt tgtcaactat ccaaaagata
      541 acaaaagaaa aatggatgag acagatgctt catcagcagt gaaagtgaaa agagcagtcc
      601 agaaaacatc cgatttaata gtgttgggtc tcccatggaa aacaaccgaa caggacctga
      661 aagagtattt tagtaccttt ggagaagttc ttatggtgca ggtcaagaaa gatcttaaga
      721 ctggtcattc aaaggggttt ggctttgttc gttttacgga atatgaaaca caagtgaaag
      781 taatgtcaca gcgacatatg atagatggac gatggtgtga ctgcaaactt cctaattcta
      841 agcaaagcca agatgagcct ttgagaagca gaaaagtgtt tgtggggcgc tgtacagagg
      901 acatgactga ggatgagctg cgggagttct tctctcagta cggggatgtg atggatgtct
      961 tcatccccaa gccattcagg gcctttgcct ttgttacatt tgcagatgat cagattgcgc
     1021 agtctctttg tggagaggac ttgatcatta aaggaatcag cgttcatata tccaatgccg
     1081 aacctaagca caatagcaat agacagttag aaagaagtgg aagatttggt ggtaatccag
     1141 gtggctttgg gaatcagggt ggatttggta atagcagagg gggtggagct ggtttgggaa
     1201 acaatcaagg tagtaatatg ggtggtggga tgaactttgg tgcgttcagc attaatccag
     1261 ccatgatggc tgccgcccag gcagcactac agagcagttg gggtatgatg ggcatgttag
     1321 ccagccagca gaaccagtca ggcccatcgg gtaataacca aaaccaaggc aacatgcaga
     1381 gggagccaaa ccaggccttc ggttctggaa ataactctta tagtggctct aattctggtg
     1441 cagcaattgg ttggggatca gcatccaatg cagggtcggg cagtggtttt aatggaggct
     1501 ttggctcaag catggattct aagtcttctg gctggggaat gaagcttatc gatagcaagg
     1561 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1621 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1681 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
     1741 tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1801 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1861 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1921 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1981 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     2041 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     2101 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     2161 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2221 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagagatct atctaggacc
     2281 cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     2341 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
     2401 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     2461 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
     2521 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc
     2581 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     2641 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     2701 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     2761 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     2821 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc
     2881 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     2941 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     3001 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     3061 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     3121 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     3181 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     3241 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     3301 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     3361 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa
     3421 gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     3481 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3541 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3601 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3661 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3721 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3781 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3841 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3901 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3961 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4021 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4081 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4141 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4201 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4261 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     4321 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     4381 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     4441 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     4501 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgg
//
