LOCUS       sequence-373718-        5000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses His8-ETV1_DBD(332-458) with a TEV site for removal of the 
            His8 tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5000)
  AUTHORS   Selig EE, Sohn EJ, Stoja A, Moreno-Romero AK, Akula S, Xu X, Bishop 
            AJR, Libich DS
  TITLE     Phase separation of the oncogenic fusion protein EWS::FLI1 is 
            modulated by its DNA-binding domain.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 May 20;122(20):e2221823122. doi: 
            10.1073/pnas.2221823122. Epub 2025 May 16.
  PUBMED    40377985
REFERENCE   2  (bases 1 to 5000)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2025-05-20"; volume: "122(20):e2221823122. 
            doi"; pages: " 10.1073/pnas.2221823122. Epub 2025 May 16"
COMMENT     Sequence Label: pAG8Ha-ETV1_DBD(332-458)
FEATURES             Location/Qualifiers
     source          1..5000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..33,34..39)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(214..261)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     257..275
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(716..736)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(743..766)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     RBS             781..803
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(834..853)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(835..853)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2555..2746
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2740..2762)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2848..2987
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2903..2922
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3002..3019)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3173..3761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3253..3272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3932..4723,4724..4792))
                     /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     4555..4574
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4793..4897)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4965..4983
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatccgata agctttaatg cggtagttta tcacagttaa
       61 attgctaacg cagtcaggca ccgtgtatga aatctaacaa tgcgctcatc gtcatcctcg
      121 gcaccgtcac cctggatgct gtaggcatag gcttggttat gccggtactg ccgggcctct
      181 tgcgggatat ccggatatag ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga
      241 ggccccaagg ggttatgcta gttattgctc agcggtggca gcagccaact cagcttcctt
      301 tcgggctttg ttagcagccg gatccttatt acgccatgct ttcatcaaag tggctcagcg
      361 gaacggtgtc ttcctcgtta atgtgacgct ccatatcggt tttcagcagc ggacgctggt
      421 tgtccggaaa cgccatgctg aacagcgcct ccggatcgca aacgaacttg tacacataac
      481 gttcgcccgc cactttttgc atgataccct tctcgtaata gtaacgcagg ctacggctca
      541 gtttgtcata gttcatcgcc ggacggttct tctgaatgcc ccaacgacgc gcaacttcct
      601 ccggttcgat cagcttaaac tccatgccac gaccggtcca cgcgatgaag tggctgttgc
      661 tcggatcgtc cagcagcgcc accaggaatt gccacagctg caggctacca cgggcgccct
      721 gaaaataaag attctcgcta gcatggtgat ggtggtggtg atggtggtcc atatggtata
      781 tctccttctt aaagttaaac aaaattattt ctagagggaa accgttgtgg tctccctata
      841 gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct ctacgccgga cgcatcgtgg
      901 ccggcatcac cggcgctaca ggtgcggttg cttgcgccta tatcgccgac ccggcatcac
      961 cggcgctaca ggtgcggttg cttgcgccta tatcgccgac atcaccgatg gggaagatcg
     1021 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     1081 ggccgtggga ctgttgggct ccatctcctt gcatggggac tgttgggctc catctccttg
     1141 catgggggga ctgttgggct ccatctcctt gcatgcacca ttccttgcgg cggcggtgct
     1201 caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg
     1261 tcaaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     1321 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     1381 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg
     1441 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc
     1501 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct
     1561 gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct
     1621 tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga
     1681 tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat
     1741 cattggaccg ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt
     1801 ggcatggatt gtaggtgccg ccctatacct tgtcatggat tgtaggtgcc gccctatacc
     1861 ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg ggccacctcg acctgaatgg
     1921 aagccggcgg cacctcgcta acggattcac cactccaaga attggagcca atcaattctt
     1981 gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac atatccatcg cgtccgccat
     2041 ctccagcagc cgcacgcggc gcatctcggg cagcgttggg tcctggccac gggtgtgcat
     2101 gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca
     2161 gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac
     2221 ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa
     2281 gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg
     2341 tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg
     2401 gtcccgccgc atccataccg ccagttgttt accctcacga cgttccagta accgggcatg
     2461 ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc
     2521 catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct
     2581 taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct
     2641 ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta
     2701 ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     2761 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     2821 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     2881 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccattgc
     2941 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     3001 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3061 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3121 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3181 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3241 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3301 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3361 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3421 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3481 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3541 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3601 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3661 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3721 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3781 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3841 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3901 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3961 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4021 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4081 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4141 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4201 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg
     4261 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4321 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4381 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4441 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4501 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     4561 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4621 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4681 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4741 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4801 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4861 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4921 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     4981 ggccctttcg tcttcaagaa
//
