LOCUS       sequence-473632-        7294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of FBXO31 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7294)
  AUTHORS   Duan S, Moro L, Qu R, Simoneschi D, Cho H, Jiang S, Zhao H, Chang Q,
            de Stanchina E, Arbini AA, Pagano M
  TITLE     Loss of FBXO31-mediated degradation of DUSP6 dysregulates ERK and 
            PI3K-AKT signaling and promotes prostate tumorigenesis.
  JOURNAL   Cell Rep. 2021 Oct 19;37(3):109870. doi: 
            10.1016/j.celrep.2021.109870.
  PUBMED    34686346
REFERENCE   2  (bases 1 to 7294)
  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: "Cell Rep.";
            date: "2021-10-19"; volume: "37(3):109870. doi"; pages: " 
            10.1016/j.celrep.2021.109870"
COMMENT     Sequence Label: pcDNA3 FLAG FBXO31
FEATURES             Location/Qualifiers
     source          1..7294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1186..1203)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1192..1416
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1462..1890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1549..1568)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1759..1780
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1899..1919)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1904..2233
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2084..2219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2146..2165
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2300..3094
                     /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(2354..2373)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2964..2983
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3268..3389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3305..3324)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3359..3378
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3438..3454)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3438..3454)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3451..3473)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3462..3478
                     /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(3486..3492,3493..3510,3511..3516))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3531..3552
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3669..3686)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3840..4428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3920..3939)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4599..5390,5391..5459))
                     /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     5222..5241
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5460..5564)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5639..5658)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5830..6209
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6210..6413
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6364..6384
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6458..6477
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6458..6476
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6518..6541
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6554..6577
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6590..6616
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             6590..6616
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6626..6652
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             6626..6652
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6719..6745
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 gaccctatga gattcaagcc tctgttcagg atccatctga tggagaggaa ggctgccaca
       61 gtggagtgca tgtacggcca caaagggccc caccacggcc acatccagat tgtgaagaag
      121 gatgagttct ccaccaagtg caaccagacg gaccaccaca ggatgtccgg cgggaggcag
      181 gaggagtttc ggacgtggct gagggaggaa tgggggcgca cgctggagga catcttccac
      241 gagcacatgc aggagctcat cctgatgaag ttcatctaca ccagtcagta cgacaactgc
      301 ctgacctacc gccgcatcta cctgccgccc agccgccccg acgacctcat caagcctggc
      361 ctcttcaaag gtacctatgg cagccacggc ctggagattg tgatgctcag cttccacggc
      421 cggcgtgcca ggggcaccaa gatcacgggc gaccccaaca tccccgctgg gcagcagaca
      481 gtggagatcg acctgaggca tcggatccag ctgcccgacc tcgagaacca gcgcaacttc
      541 aatgagctct cccgcatcgt cctggaggtg cgcgagaggg tgcgccagga gcagcaggaa
      601 ggcgggcacg aggcgggcga gggtcgtggc cggcagggcc cccgggagtc ccagccaagc
      661 cctgcccagc ccagggcaga ggcgcccagc aagggcccag atgggacacc tggtgaggat
      721 ggtggcgagc ctggggatgc cgtagctgcg gccgagcagc ctgcccagtg tgggcagggg
      781 cagccgttcg tgctgcccgt gggcgtgagc tccaggaatg aggactaccc ccgaacctgc
      841 aggatgtgtt tttatggcac aggcctcatc gcgggccacg gcttcaccag ccctgaacgc
      901 acccccgggg tcttcatcct cttcgatgag gaccgcttcg ggttcgtctg gctggagctg
      961 aaatccttca gcctgtacag ccgggtccag gccaccttcc ggaacgcaga tgcgccgtcc
     1021 ccacaggcct tcgatgagat gctcaagaac attcagtccc tcacctcctg aatcgattgg
     1081 cgccttaatt aaggcctagg cggccgcggc gcgccgaatt ctgcagatat ccagcacagt
     1141 ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     1201 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1261 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     1321 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     1381 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     1441 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1501 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     1561 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     1621 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     1681 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     1741 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     1801 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     1861 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     1921 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     1981 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2041 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2101 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2161 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2221 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     2281 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     2341 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     2401 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     2461 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     2521 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     2581 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     2641 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     2701 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     2761 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     2821 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     2881 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     2941 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3001 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3061 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     3121 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     3181 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     3241 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     3301 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3361 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     3421 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3481 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3541 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3601 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3661 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3721 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3781 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3841 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3901 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3961 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4021 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4081 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4141 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4201 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4261 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4321 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4381 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4441 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4501 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4561 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4621 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4681 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4741 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4801 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4861 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4921 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4981 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5041 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5101 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5161 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5221 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5281 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5341 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5401 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5461 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5521 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5581 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     5641 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     5701 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     5761 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     5821 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     5881 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     5941 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     6001 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     6061 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     6121 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     6181 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     6241 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     6301 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     6361 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
     6421 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
     6481 cccaagctgg ctagcgttta aacttaagct taccatggac tacaaggacg acgatgacaa
     6541 ggtaccagga tctgactaca aggacgacga tgacaagctc gatggaggat acccctacga
     6601 cgtgcccgac tacgccgctg gagcataccc ctacgacgtg cccgactacg cccaaaactt
     6661 atacttccaa ggacaaaact tatacttcca aggaggatcc agtgtggtgg aattcatgta
     6721 cccatacgat gttccagatt acgctatggc ggtgtgtgct cgcctttgcg gcgtgggccc
     6781 gtcgcgcgga tgtcggcgcc gccagcagcg ccggggcccg gccgagacgg cggcggccga
     6841 cagcgagccg gacacagacc ccgaggagga gcgcatcgag gctagcgccg gggtcggggg
     6901 cggcttgtgc gcgggcccct cgccgccgcc cccgcgctgc tcgctgctgg agctgccgcc
     6961 cgagctgctg gtggagatct tcgcgtcgct gccgggcacg gacctaccca gcttggccca
     7021 ggtctgcacg aagttccggc gcatcctcca caccgacacc atctggagga ggcgttgccg
     7081 tgaggagtat ggtgtttgcg aaaacttgcg gaagctggag atcacaggcg tgtcttgtcg
     7141 ggacgtctat gcgaagctgc ttcaccgata tagacacatt ttgggattgt ggcagccaga
     7201 tatcgggcca tacggaggac tgctgaacgt ggtggtggac ggcctgttca tcatcgggtg
     7261 gatgtacctg cctccccatg acccccacgt cgat
//
