LOCUS       sequence-390439-        5464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5464)
  AUTHORS   Bouchard JJ, Otero JH, Scott DC, Szulc E, Martin EW, Sabri N, 
            Granata D, Marzahn MR, Lindorff-Larsen K, Salvatella X, Schulman BA,
            Mittag T
  TITLE     Cancer Mutations of the Tumor Suppressor SPOP Disrupt the Formation 
            of Active, Phase-Separated Compartments.
  JOURNAL   Mol Cell. 2018 Oct 4;72(1):19-36.e8. doi: 
            10.1016/j.molcel.2018.08.027. Epub 2018 Sep 20.
  PUBMED    30244836
REFERENCE   2  (bases 1 to 5464)
  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: 
            "10.1016/j.molcel.2018.08.027"; journalName: "Mol Cell"; date: 
            "2018-10-4- 4"; volume: "72"; issue: "1"; pages: "19-36.e8"
COMMENT     Sequence Label: HisTEV-cDAXX
FEATURES             Location/Qualifiers
     source          1..5464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             113..130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    140..164
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             167..187
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(933..957)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1010..1028)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1024..1071
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1437..1442,1443..1459,1460..1465))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1492..1510)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1578..1682
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1683..1751,1752..2543)
                     /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(1901..1920)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2714..3302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3203..3222
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3456..3473
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3488..3627
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3553..3572)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3713..3735
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3729..3920)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5382..5401
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacat atgtcgtact accatcacca
      121 tcaccatcac ctcgaatcaa caagtttgta caaaaaagca ggcttcgaaa acctgtattt
      181 tcagggcgcc agccccatgt cctcactaca gatctccaat gaaaagaacc tggaacctgg
      241 caaacagatc agcagatctt caggggagca gcaaaacaaa ggacgcatag tgtcaccatc
      301 gttactgtca gaagaacccc tggccccctc cagcatagat gctgaaagca atggagaaca
      361 gcctgaggag ctgaccctgg aggaagaaag ccctgtgtct cagctctttg agctagagat
      421 tgaagctttg cctctggata ccccttcctc tgtggagacg gacatttcct cttccaggaa
      481 gcaatcagag gagcccttca ccactgtctt agagaatgga gcaggcatgg tctcttctac
      541 ttccttcaat ggaggcgtct ctcctcacaa ctggggagat tctggtcccc cctgcaaaaa
      601 atctcggaag gagaagaagc aaacaggatc agggccatta ggaaacagct atgtggaaag
      661 gcaaaggtca gtgcatgaga agaatgggaa aaagatatgt accctgccca gcccaccttc
      721 ccccttggct tccttggccc cagttgctga ttcctccacg agggtggact ctcccagcca
      781 tggcctggtg accagctccc tctgcatccc ttctccagcc cggctgtccc aaacccccca
      841 ttcacagcct cctcggcctg gtacttgcaa gacaagtgtg gccacacaat gcgatccaga
      901 agagatcatc gtgctctcag actctgatta ggacccagct ttcttgtaca aagtggttga
      961 ttcgaggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     1021 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     1081 ggaactatat ccggatatcc acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt
     1141 ggctccaagt agcgaagcga gcaggactgg gcggcggcca aagcggtcgg acagtgctcc
     1201 gagaacgggt gcgcatagaa attgcatcaa cgcatatagc gctagcagca cgccatagtg
     1261 actggcgatg ctgtcggaat ggacgatatc ccgcaagagg cccggcagta ccggcataac
     1321 caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt
     1381 agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta
     1441 aagcttatcg atgataagct gtcaaacatg agaattcttg aagacgaaag ggcctcgtga
     1501 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     1561 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1621 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1681 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1741 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1801 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1861 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1921 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1981 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2041 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2101 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2161 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2221 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2281 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2341 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2401 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2461 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2521 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2581 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2641 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2701 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2761 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2821 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     2881 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2941 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3001 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3061 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3121 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3181 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3241 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3301 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3361 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3421 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3481 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
     3541 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     3601 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     3661 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3721 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     3781 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     3841 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     3901 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     3961 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     4021 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     4081 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     4141 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     4201 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     4261 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     4321 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
     4381 acccgtggcc aggacccaac gctgcccgag atgcgccgcg tgcggctgct ggagatggcg
     4441 gacgcgatgg atatgttctg ccaagggttg gtttgcgcat tcacagttct ccgcaagaat
     4501 tgattggctc caattcttgg agtggtgaat ccgttagcga ggtgccgccg gcttccattc
     4561 aggtcgaggt ggcccggctc catgcaccgc gacgcaacgc ggggaggcag acaaggtata
     4621 gggcggcgcc tacaatccat gccaacccgt tccatgtgct cgccgaggcg gcataaatcg
     4681 ccgtgacgat cagcggtcca atgatcgaag ttaggctggt aagagccgcg agcgatcctt
     4741 gaagctgtcc ctgatggtcg tcatctacct gcctggacag catggcctgc aacgcgggca
     4801 tcccgatgcc gccggaagcg agaagaatca taatggggaa ggccatccag cctcgcgtcg
     4861 cgaacgccag caagacgtag cccagcgcgt cggccgccat gccggcgata atggcctgct
     4921 tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga
     4981 ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc
     5041 cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag
     5101 tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     5161 aggctctcaa gggcatcggt cgatcgacgc tctcccttat gcgactcctg cattaggaag
     5221 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     5281 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     5341 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     5401 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     5461 atcg
//
