LOCUS       sequence-334855-        6578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of mouse SPOP Y353E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6578)
  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 6578)
  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: SPOP-V5 (BACK))
FEATURES             Location/Qualifiers
     source          1..6578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(114..134)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        119..448
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      299..434
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     361..380
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             515..1309
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(569..588)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1179..1198
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1483..1604
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1520..1539)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1574..1593
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1653..1669)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1653..1669)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1666..1688)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1677..1693
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1701..1707,1708..1725,1726..1731))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1746..1767
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1884..1901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2055..2643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2135..2154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2814..3605,3606..3674))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3437..3456
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3675..3779)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3854..3873)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4045..4424
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4425..4628
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4579..4599
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4673..4692
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4673..4691
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4781..4822
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(5979..5996)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5985..6209
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6255..6578,1..105)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6342..6361)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6552..6573
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
       61 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
      121 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
      181 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
      241 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
      301 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
      361 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
      421 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
      481 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
      541 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
      601 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
      661 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
      721 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
      781 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
      841 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
      901 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
      961 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     1021 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     1081 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     1141 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     1201 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     1261 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     1321 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     1381 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     1441 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     1501 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     1561 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     1621 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1681 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1741 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1801 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1861 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1921 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1981 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2041 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2101 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2161 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2221 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2281 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2341 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2401 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2461 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2521 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2581 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2641 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2701 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2761 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2821 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2881 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2941 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3001 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3061 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3121 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3181 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3241 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3301 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3361 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3421 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3481 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3541 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3601 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3661 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3721 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3781 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     3841 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     3901 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     3961 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
     4021 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
     4081 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     4141 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     4201 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     4261 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     4321 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     4381 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     4441 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     4501 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     4561 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     4621 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga
     4681 ctcactatag ggagacccaa gctggctagc gtttaaactt aagcttggta ccgagctcgg
     4741 atccactagt ccagtgtggt ggaattctgc agatatcatg ggtaagccta tccctaaccc
     4801 tctcctcggt ctcgattcta cgatgtcaag ggttccaagt cctccacctc cggcagaaat
     4861 gtcgagtggt cccgttgccg agagctggtg ctatacacag atcaaggtag tgaagttctc
     4921 ctacatgtgg accatcaata actttagttt ttgccgggag gaaatgggtg aagtcataaa
     4981 aagttcaact ttttcatcag gagccaatga taaactgaaa tggtgtttgc gagtaaaccc
     5041 gaaagggcta gatgaagaaa gcaaagatta cctgtcgctt tacctgttgt tggtcagctg
     5101 tccaaagagt gaagttcggg caaagttcaa gttctccatc ctgaatgcca agggagaaga
     5161 aaccaaagct atggagagtc agcgagctta taggttcgtg caaggcaaag actggggatt
     5221 caaaaaattc atccgcagag attttctctt ggacgaggcc aatgggcttc tccctgatga
     5281 caagctcacc ctcttctgtg aggtgagtgt agtacaagac tctgtcaata tttctgggca
     5341 gaatactatg aacatggtaa aggttcctga atgccggttg gcagatgagt taggagggct
     5401 gtgggagaac tctcgattca ccgactgctg cttgtgtgtg gctggtcaag aattccaggc
     5461 tcacaaggct atcttagcag ctcggtctcc agtcttcagt gccatgtttg aacatgaaat
     5521 ggaggagagc aaaaagaatc gggttgagat taacgacgtg gagcctgagg tgtttaagga
     5581 gatgatgtgc ttcatctaca cggggaaggc gccgaacctc gacaagatgg ctgacgactt
     5641 gctggcagct gccgataagt acgccctgga gcgtctgaag gtcatgtgtg aggacgccct
     5701 ctgcagtaac ctctctgtgg agaacgctgc agagattctc atcctggctg acctccacag
     5761 cgcagatcag ctgaagacgc aggcagttga tttcatcaac tatcatgctt cggatgtctt
     5821 ggagacctct gggtggaagt caatggtggt gtcacatccc cacttagtgg cagaagcaga
     5881 gcgttctctg gcttcagcac agtgcccttt cctggggccc ccacgcaaac gcctgaagca
     5941 atcctaatct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     6001 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     6061 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     6121 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     6181 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     6241 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     6301 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     6361 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6421 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6481 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6541 gttctttaat agtggactct tgttccaaac tggaacaa
//
