LOCUS       sequence-359561-        5405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the construct His8-EWS_1-264 with a TEV cleavage site for 
            the removal of the His8 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5405)
  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 5405)
  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-EWS_1-264
FEATURES             Location/Qualifiers
     source          1..5405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3..21
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             53..75
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             90..113
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             120..140
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(986..1004)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1000..1047
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1222..1227,1228..1245,1246..1251))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1278..1296)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1364..1468
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1469..1537,1538..2329)
                     /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(1687..1706)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2500..3088
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2989..3008
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3242..3259
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3274..3413
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3339..3358)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3499..3521
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3515..3706)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata attttgttta
       61 actttaagaa ggagatatac catatggacc accatcacca ccaccatcac catgctagcg
      121 agaatcttta ttttcagggc gccgcgagca ccgattatag cacctatagc caggccgcgg
      181 cccagcaggg ttatagcgcc tataccgcgc agccgaccca gggttatgcc cagaccaccc
      241 aggcgtatgg ccagcagagc tatggtacct atggccagcc gaccgatgtg agctataccc
      301 aggcccagac caccgcgacc tatggtcaga ccgcctatgc caccagctat ggtcagccgc
      361 cgaccggtta taccaccccg accgccccgc aggcctatag ccagccggtt cagggctatg
      421 gtaccggtgc ctatgatacc accaccgcca ccgtgaccac cacccaggca agctatgcgg
      481 cccagagcgc ctatggtacc cagccggcat atccggccta tggtcagcag ccggcggcca
      541 ccgcaccgac ccgcccgcag gatggtaaca aaccgaccga aaccagccag ccgcagagca
      601 gcaccggcgg ttataaccag ccgagcctgg gttatggcca gagcaattat agctatccgc
      661 aggttccggg cagctatccg atgcagccgg ttaccgcacc gccgagctat ccgccgacca
      721 gctatagcag cacccagccg accagctatg atcagagcag ctatagccag cagaacacct
      781 atggtcagcc gagcagctat ggccagcaga gcagctatgg tcaacaaagc agctatggtc
      841 agcagccgcc gaccagctat ccgccgcaga ccggtagcta tagccaggct ccgagccagt
      901 atagccagca gagctcttct tatggccagc agtgaggatc cggctgctaa caaagcccga
      961 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     1021 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atatcccgca
     1081 agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc
     1141 cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat
     1201 ttaactgtga taaactaccg cattaaagct tatcggatga taagctgtca aacatgagaa
     1261 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     1321 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1381 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1441 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1501 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1561 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1621 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     1681 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     1741 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1801 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1861 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1921 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1981 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     2041 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2101 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2161 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2221 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2281 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2341 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     2401 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2461 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2521 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2581 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2641 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2701 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2761 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2821 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2881 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2941 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3001 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3061 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3121 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3181 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3241 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     3301 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     3361 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     3421 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3481 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3541 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     3601 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     3661 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     3721 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     3781 tacgggttac tgatgatgaa catgcccggt tactggaacg tcgtgagggt aaacaactgg
     3841 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     3901 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     3961 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     4021 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     4081 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     4141 tcaacgacag gagcacgatc atgcacaccc gtggccagga cccaacgctg cccgagatgc
     4201 gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt
     4261 gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt
     4321 tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg
     4381 caacgcgggg aggcagacaa ggtatagggc ggcacctaca atccatgaca aggtataggg
     4441 cggcacctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg
     4501 tgacgatcag cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa
     4561 gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc
     4621 cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga
     4681 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct
     4741 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     4801 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     4861 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     4921 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     4981 ctctcaaggg catcggttga cgctctccct tatgcgactc ctgcattagg aagcagccca
     5041 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     5101 agcccaacag tccccccatg caaggagatg gagcccaaca gtccccatgc aaggagatgg
     5161 agcccaacag tcccacggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     5221 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgcaag
     5281 caaccgcacc tgtagcgccg gtgatgccgg gtcggcgata taggcgcaag caaccgcacc
     5341 tgtagcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
     5401 gcgaa
//
