LOCUS       sequence-386369-        5189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the RRM-RG2-ZnF (359-549) of EWS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5189)
  AUTHORS   Selig EE, Bhura R, White MR, Akula S, Hoffman RD, Tovar CN, Xu X, 
            Booth RE, Libich DS
  TITLE     Biochemical and biophysical characterization of the nucleic acid 
            binding properties of the RNA/DNA binding protein EWS.
  JOURNAL   Biopolymers. 2023 May;114(5):e23536. doi: 10.1002/bip.23536. Epub 
            2023 Mar 17.
  PUBMED    36929870
REFERENCE   2  (bases 1 to 5189)
  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: 
            "Biopolymers."; date: "2023-05"; volume: "114(5):e23536. doi"; 
            pages: " 10.1002/bip.23536. Epub 2023 Mar 17"
COMMENT     Sequence Label: pAG8Ha-His-EWSR1-359-549
FEATURES             Location/Qualifiers
     source          1..5189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(206..224)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      220..267
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(442..447,448..465,466..471))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(498..516)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        584..688
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(689..757,758..1549)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(907..926)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1720..2308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2209..2228
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2462..2479
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2494..2633
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2559..2578)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2719..2741
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2735..2926)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4628..4647
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4628..4646
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4678..4700
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4715..4738
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             4745..4765
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 cgcgtggtcc gcgtggcagc cgtggtaacc cgagcggcgg tggcaacgtt cagcaccgtg
       61 cgggtgattg gcaatgcccg aacccgggtt gcggcaacca gaacttcgcg tggcgtaccg
      121 agtgcaacca atgcaaggcg ccgaaaccgg aatgaggatc cggctgctaa caaagcccga
      181 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
      241 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atatcccgca
      301 agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc
      361 cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat
      421 ttaactgtga taaactaccg cattaaagct tatcggatga taagctgtca aacatgagaa
      481 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
      541 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      661 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      721 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      781 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      841 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      901 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      961 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1021 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1081 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1141 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1201 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     1261 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1321 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1381 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1441 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1501 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1561 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1621 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1861 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1921 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1981 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2041 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2101 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2161 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2221 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2281 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2341 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2401 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2461 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2521 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     2581 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     2641 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     2701 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     2761 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     2821 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     2881 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     2941 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     3001 tacgggttac tgatgatgaa catgcccggt tactggaacg tcgtgagggt aaacaactgg
     3061 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     3121 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     3181 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     3241 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     3301 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     3361 tcaacgacag gagcacgatc atgcacaccc gtggccagga cccaacgctg cccgagatgc
     3421 gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt
     3481 gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt
     3541 tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg
     3601 caacgcgggg aggcagacaa ggtatagggc ggcacctaca atccatgaca aggtataggg
     3661 cggcacctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg
     3721 tgacgatcag cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa
     3781 gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc
     3841 cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga
     3901 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct
     3961 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     4021 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     4081 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     4141 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     4201 ctctcaaggg catcggttga cgctctccct tatgcgactc ctgcattagg aagcagccca
     4261 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     4321 agcccaacag tccccccatg caaggagatg gagcccaaca gtccccatgc aaggagatgg
     4381 agcccaacag tcccacggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     4441 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgcaag
     4501 caaccgcacc tgtagcgccg gtgatgccgg gtcggcgata taggcgcaag caaccgcacc
     4561 tgtagcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
     4621 gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag aaataatttt
     4681 gtttaacttt aagaaggaga tataccatat ggaccaccat caccaccacc atcaccatgc
     4741 tagcgagaat ctttattttc agggcgccga taacagcgcg atctacgtgc agggtctgaa
     4801 cgacagcgtt accctggacg atctggcgga tttctttaag cagtgcggcg tggttaagat
     4861 gaacaaacgt accggtcaac cgatgatcca catttacctg gataaagaaa ccggcaagcc
     4921 gaaaggtgac gcgaccgtga gctacgagga cccgccgacc gcgaaggcgg cggttgaatg
     4981 gttcgacggc aaagattttc agggtagcaa gctgaaagtg agcctggcgc gtaagaaacc
     5041 gccgatgaac agcatgcgtg gcggtctgcc gccgcgtgag ggtcgtggca tgccgccgcc
     5101 actgcgtggc ggtccgggcg gtccgggcgg tccgggcggt ccgatgggtc gtatgggcgg
     5161 tcgtggcggt gaccgtggcg gttttccgc
//
