LOCUS       sequence-163027-        4530 bp    DNA     circular SYN 14-APR-2025
DEFINITION  DONOR vector for Gateway cloning of EWSR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4530)
  AUTHORS   Couthouis J, Hart MP, Erion R, King OD, Diaz Z, Nakaya T, Ibrahim F,
            Kim HJ, Mojsilovic-Petrovic J, Panossian S, Kim CE, Frackelton EC, 
            Solski JA, Williams KL, Clay-Falcone D, Elman L, McCluskey L, Greene
            R, Hakonarson H, Kalb RG, Lee VM, Trojanowski JQ, Nicholson GA, 
            Blair IP, Bonini NM, Van Deerlin VM, Mourelatos Z, Shorter J, Gitler
            AD
  TITLE     Evaluating the role of the FUS/TLS-related gene EWSR1 in amyotrophic
            lateral sclerosis.
  JOURNAL   Hum Mol Genet. 2012 Jul 1;21(13):2899-911. doi: 10.1093/hmg/dds116. 
            Epub 2012 Mar 27.
  PUBMED    22454397
REFERENCE   2  (bases 1 to 4530)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1093/hmg/dds116"; journalName: "Hum Mol Genet"; date: 
            "2012-07-1- 1"; volume: "21"; issue: "13"; pages: "2899-911"
COMMENT     Sequence Label: pDONR221-EWSR1 Stop
FEATURES             Location/Qualifiers
     source          1..4530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2655..2754)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2771..2790)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2772..2790)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2795..2811)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2795..2811)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2913..2932)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2924..3733
                     /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(2991..3010)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3880..4468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4369..4388
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttc gaaggagata acaaaatggc gtccacggat tacagtacct atagccaagc
      721 tgcagcgcag cagggctaca gtgcttacac cgcccagccc actcaaggat atgcacagac
      781 cacccaggca tatgggcaac aaagctatgg aacctatgga cagcccactg atgtcagcta
      841 tacccaggct cagaccactg caacctatgg gcagaccgcc tatgcaactt cttatggaca
      901 gcctcccact ggttatacta ctccaactgc cccccaggca tacagccagc ctgtccaggg
      961 gtatggcact ggtgcttatg ataccaccac tgctacagtc accaccaccc aggcctccta
     1021 tgcagctcag tctgcatatg gcactcagcc tgcttatcca gcctatgggc agcagccagc
     1081 agccactgca cctacaagac cgcaggatgg aaacaagccc actgagacta gtcaacctca
     1141 atctagcaca gggggttaca accagcccag cctaggatat ggacagagta actacagtta
     1201 tccccaggta cctgggagct accccatgca gccagtcact gcacctccat cctaccctcc
     1261 taccagctat tcctctacac agccgactag ttatgatcag agcagttact ctcagcagaa
     1321 cacctatggg caaccgagca gctatggaca gcagagtagc tatggtcaac aaagcagcta
     1381 tgggcagcag cctcccacta gttacccacc ccaaactgga tcctacagcc aagctccaag
     1441 tcaatatagc caacagagca gcagctacgg gcagcagagt tcattccgac aggaccaccc
     1501 cagtagcatg ggtgtttatg ggcaggagtc tggaggattt tccggaccag gagagaaccg
     1561 gagcatgagt ggccctgata accggggcag gggaagaggg ggatttgatc gtggaggcat
     1621 gagcagaggt gggcggggag gaggacgcgg tggaatgggc gctggagagc gaggtggctt
     1681 caataagcct ggtggaccca tggatgaagg accagatctt gatctaggcc cacctgtaga
     1741 tccagatgaa gactctgaca acagtgcaat ttatgtacaa ggattaaatg acagtgtgac
     1801 tctagatgat ctggcagact tctttaagca gtgtggggtt gttaagatga acaagagaac
     1861 tgggcaaccc atgatccaca tctacctgga caaggaaaca ggaaagccca aaggcgatgc
     1921 cacagtgtcc tatgaagacc cacccactgc caaggctgcc gtggaatggt ttgatgggaa
     1981 agattttcaa gggagcaaac ttaaagtctc ccttgctcgg aagaagcctc caatgaacag
     2041 tatgcggggt ggtctgccac cccgtgaggg cagaggcatg ccaccaccac tccgtggagg
     2101 tccaggaggc ccaggaggtc ctgggggacc catgggtcgc atgggaggcc gtggaggaga
     2161 tagaggaggc ttccctccaa gaggaccccg gggttcccga gggaacccct ctggaggagg
     2221 aaacgtccag caccgagctg gagactggca gtgtcccaat ccgggttgtg gaaaccagaa
     2281 cttcgcctgg agaacagagt gcaaccagtg taaggcccca aagcctgaag gcttcctccc
     2341 gccacccttt ccgcccccgg gtggtgatcg tggcagaggt ggccctggtg gcatgcgggg
     2401 aggaagaggt ggcctcatgg atcgtggtgg tcccggtgga atgttcagag gtggccgtgg
     2461 tggagacaga ggtggcttcc gtggtggccg gggcatggac cgaggtggct ttggtggagg
     2521 aagacgaggt ggccctgggg ggccccctgg acctttgatg gaacagatgg gaggaagaag
     2581 aggaggacgt ggaggacctg gaaaaatgga taaaggcgag caccgtcagg agcgcagaga
     2641 tcggccctac taggacccag ctttcttgta caaagttggc attataagaa agcattgctt
     2701 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2761 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2821 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaataatat catcatgaac
     2881 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2941 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     3001 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     3061 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     3121 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     3181 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     3241 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     3301 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     3361 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     3421 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     3481 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3541 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3601 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3661 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3721 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3781 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3841 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3901 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3961 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4021 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4081 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4141 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4201 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4261 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4321 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4381 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4441 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4501 ggccttttgc tggccttttg ctcacatgtt
//
