LOCUS       sequence-359563-        5219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the construct His8-sumo-EWS_91-199.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5219)
  AUTHORS   Johnson CN, Xu X, Holloway SP, Libich DS
  TITLE     The (1)H, (15)N and (13)C resonance assignments of the 
            low-complexity domain from the oncogenic fusion protein EWS-FLI1.
  JOURNAL   Biomol NMR Assign. 2022 Apr;16(1):67-73. doi: 
            10.1007/s12104-021-10061-4. Epub 2022 Jan 7.
  PUBMED    34994941
REFERENCE   2  (bases 1 to 5219)
  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: 
            "10.1007/s12104-021-10061-4"; journalName: "Biomol NMR Assign"; 
            date: "2022-04"; volume: "16"; issue: "1"; pages: "67-73"
COMMENT     Sequence Label: pAG8H-sumo-EWS_91-199
FEATURES             Location/Qualifiers
     source          1..5219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     56..75
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        56..74
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             106..128
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             143..166
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(853..871)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      867..914
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1089..1094,1095..1112,1113..1118))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1145..1163)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1231..1335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1336..1404,1405..2196)
                     /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(1554..1573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2367..2955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2856..2875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3109..3126
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3141..3280
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3206..3225)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3366..3388
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3382..3573)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata
       61 cgactcacta tagggagacc acaacggttt ccctctagaa ataattttgt ttaactttaa
      121 gaaggagata taccatatgg accaccatca ccaccaccat caccatgcta gcggatctgc
      181 tgcgggtggc gaagaagata agaaaccggc aggtggcgaa ggtggcggtg cccatatcaa
      241 cctgaaagtg aaaggtcaag acggcaacga agtctttttc cgcatcaaac gttctaccca
      301 gctgaaaaag ctgatgaacg catactgtga ccgtcagtct gtagacatga ccgcaattgc
      361 tttcctgttt gatggtcgtc gcctgcgtgc ggaacagacc ccggatgaac tggagatgga
      421 agatggcgac gaaatcgacg caatgctgca ccagactggt ggcgcccagg gctatggtac
      481 cggtgcctat gataccacca ccgccaccgt gaccaccacc caggcaagct atgcggccca
      541 gagcgcctat ggtacccagc cggcatatcc ggcctatggt cagcagccgg cggccaccgc
      601 accgacccgc ccgcaggatg gtaacaaacc gaccgaaacc agccagccgc agagcagcac
      661 cggcggttat aaccagccga gcctgggtta tggccagagc aattatagct atccgcaggt
      721 tccgggcagc tatccgatgc agccggttac cgcaccgccg agctatccgc cgaccagcta
      781 tagcagcacc cagtgaggtc tcggatccgg ctgctaacaa agcccgaaag gaagctgagt
      841 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
      901 tgaggggttt tttgctgaaa ggaggaacta tatccggata tcccgcaaga ggcccggcag
      961 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
     1021 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
     1081 actaccgcat taaagcttat cggatgataa gctgtcaaac atgagaattc ttgaagacga
     1141 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     1201 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1261 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1321 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1381 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1441 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1501 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1561 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     1621 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1681 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1741 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1801 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1861 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     1921 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     1981 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2041 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2101 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2161 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2221 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2281 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2341 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2401 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2461 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     2521 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2581 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2641 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2701 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2761 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2821 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2881 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2941 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3001 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3061 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3121 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3181 tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     3241 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     3301 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3361 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     3421 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     3481 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     3541 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     3601 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     3661 tgatgaacat gcccggttac tggaacgtcg tgagggtaaa caactggcgg tatggatgcg
     3721 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     3781 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     3841 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     3901 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     3961 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     4021 cacgatcatg cacacccgtg gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct
     4081 gctggagatg gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg cattcacagt
     4141 tctccgcaag aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg
     4201 ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg
     4261 cagacaaggt atagggcggc acctacaatc catgacaagg tatagggcgg cacctacaat
     4321 ccatgccaac ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg
     4381 tccaatgatc gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg
     4441 gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga
     4501 agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac
     4561 gtagcccagc gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     4621 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     4681 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     4741 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     4801 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     4861 cggttgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     4921 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggagc ccaacagtcc
     4981 ccccatgcaa ggagatggag cccaacagtc cccatgcaag gagatggagc ccaacagtcc
     5041 cacggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     5101 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgcaagcaa ccgcacctgt
     5161 agcgccggtg atgccgggtc ggcgatatag gcgcaagcaa ccgcacctgt agcgccggt
//
