LOCUS       sequence-468720-        5173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged EWSR1/ZNF384.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5173)
  TITLE     Mittag bacterial expression plasmids
REFERENCE   2  (bases 1 to 5173)
  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"
COMMENT     Sequence Label: pDest17_EWSR1/ZNF384 fusion protein LCD (1-149)
FEATURES             Location/Qualifiers
     source          1..5173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(271..276,277..293,294..299))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(326..344)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        412..516
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(517..585,586..1377)
                     /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(735..754)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1548..2136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2037..2056
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2290..2307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2322..2461
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2387..2406)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2547..2569
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2563..2754)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4216..4235
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4319..4338
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4319..4337
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4369..4391
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4411..4428
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4438..4462
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4465..4485
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4940..4964)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5017..5035)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5031..5078
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ctgggcggcg gccaaagcgg tcggacagtg ctccgagaac gggtgcgcat agaaattgca
       61 tcaacgcata tagcgctagc agcacgccat agtgactggc gatgctgtcg gaatggacga
      121 tatcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg
      181 tgacggtgcc gaggatgacg atgagcgcat tgttagattt catacacggt gcctgactgc
      241 gttagcaatt taactgtgat aaactaccgc attaaagctt atcgatgata agctgtcaaa
      301 catgagaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
      361 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
      421 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
      481 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
      541 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
      601 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
      661 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
      721 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
      781 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
      841 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
      901 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
      961 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     1021 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     1081 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     1141 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     1201 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     1261 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     1321 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     1381 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     1441 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     1501 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1561 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1621 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1681 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     1741 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     1801 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1861 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     1921 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     1981 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2041 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2101 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2161 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     2221 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     2281 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     2341 cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg
     2401 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     2461 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     2521 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     2581 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     2641 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     2701 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     2761 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     2821 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     2881 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     2941 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     3001 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     3061 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     3121 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     3181 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc
     3241 cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg
     3301 gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt
     3361 gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca
     3421 ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac
     3481 ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccaatgatc
     3541 gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct
     3601 acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga
     3661 atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc
     3721 gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     3781 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     3841 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     3901 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     3961 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgatcg
     4021 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     4081 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     4141 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     4201 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     4261 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta
     4321 atacgactca ctatagggag accacaacgg tttccctcta gaaataattt tgtttaactt
     4381 taagaaggag atatacatat gtcgtactac catcaccatc accatcacct cgaatcaaca
     4441 agtttgtaca aaaaagcagg ctccgaaaac ctgtatttcc agggtagtca gaccggtagc
     4501 tatagccagg caccgagtca gtatagtcag cagagtagta gctatggcca gcagagttca
     4561 tttcgccagg atcatccgag cagtatgggt gtttatggtc aggaaagcgg cggctttagc
     4621 ggcccgggtg aaaatcgtag catgagcggc ccggataatc gcggccgcgg tcgtggtggc
     4681 tttgatcgcg gcggcatgag tcgtggtggt cgtggcggtg gtcgtggtgg tatgggcagc
     4741 gccggcgaac gcggtggttt taataagccg ggtggcccga tggatgaagg cccggatctg
     4801 gatctggaaa gcagcggtgc actggaacgt cgtcgtaata agaccgccgg tggtcgcgaa
     4861 cgtgaaggtc gtatggaaga aagtcatttt aatagcaatc cgtatttctg gccgagcatt
     4921 ccgaccgtga gtggctaaaa cccagctttc ttgtacaaag tggttgattc gaggctgcta
     4981 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     5041 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     5101 gatatccaca ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc
     5161 gaagcgagca gga
//
