LOCUS       sequence-473495-        7150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of FBXW7gamma in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7150)
  AUTHORS   Busino L, Millman SE, Scotto L, Kyratsous CA, Basrur V, O'Connor O, 
            Hoffmann A, Elenitoba-Johnson KS, Pagano M
  TITLE     Fbxw7alpha- and GSK3-mediated degradation of p100 is a pro-survival 
            mechanism in multiple myeloma.
  JOURNAL   Nat Cell Biol. 2012 Mar 4;14(4):375-85. doi: 10.1038/ncb2463.
  PUBMED    22388891
REFERENCE   2  (bases 1 to 7150)
  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"; journalName: 
            "Nat Cell Biol"; date: "2012-03-4- 4"; volume: "14"; issue: "4"; 
            pages: "375-85"
COMMENT     Sequence Label: pcDNA3 FLAG Fbxw7gamma
FEATURES             Location/Qualifiers
     source          1..7150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     152..171
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(390..494)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(569..588)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        760..1139
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1140..1343
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1294..1314
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1388..1407
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1388..1406
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1448..1471
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3266..3283)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3272..3496
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3542..3970
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3629..3648)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3839..3860
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3979..3999)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3984..4313
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4164..4299
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4226..4245
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4380..5174
                     /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(4434..4453)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5044..5063
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5348..5469
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5385..5404)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5439..5458
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5518..5534)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5518..5534)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5531..5553)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5542..5558
                     /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(5566..5572,5573..5590,5591..5596))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5611..5632
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5749..5766)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5920..6508)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6000..6019)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6679..7150,1..320,321..389))
                     /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"
ORIGIN
        1 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
       61 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
      121 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
      181 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
      241 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
      301 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
      361 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
      421 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
      481 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
      541 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
      601 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
      661 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
      721 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
      781 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      841 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      901 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      961 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     1021 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1081 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1141 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     1201 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     1261 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     1321 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
     1381 tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta aacttaagct
     1441 taccatggac tacaaggacg acgatgacaa ggtacctagg atccagtgtg gtggaattcg
     1501 cccttgcaag cttatgggtt tctacggcac attaaaaatg attttttaca aaatgaaaag
     1561 aaagttggac catggttctg aggtccgctc tttttctttg ggaaagaaac catgcaaagt
     1621 ctcagaatat acaagtacca ctgggcttgt accatgttca gcaacaccaa caacttttgg
     1681 ggacctcaga gcagccaatg gccaagggca acaacgacgc cgaattacat ctgtccagcc
     1741 acctacaggc ctccaggaat ggctaaaaat gtttcagagc tggagtggac cagagaaatt
     1801 gcttgcttta gatgaactca ttgatagttg tgaaccaaca caagtaaaac atatgatgca
     1861 agtgatagaa ccccagtttc aacgagactt catttcattg ctccctaaag agttggcact
     1921 ctatgtgctt tcattcctgg aacccaaaga cctgctacaa gcagctcaga catgtcgcta
     1981 ctggagaatt ttggctgaag acaaccttct ctggagagag aaatgcaaag aagaggggat
     2041 tgatgaacca ttgcacatca agagaagaaa agtaataaaa ccaggtttca tacacagtcc
     2101 atggaaaagt gcatacatca gacagcacag aattgatact aactggaggc gaggagaact
     2161 caaatctcct aaggtgctga aaggacatga tgatcatgtg atcacatgct tacagttttg
     2221 tggtaaccga atagttagtg gttctgatga caacacttta aaagtttggt cagcagtcac
     2281 aggcaaatgt ctgagaacat tagtgggaca tacaggtgga gtatggtcat cacaaatgag
     2341 agacaacatc atcattagtg gatctacaga tcggacactc aaagtgtgga atgcagagac
     2401 tggagaatgt atacacacct tatatgggca tacttccact gtgcgttgta tgcatcttca
     2461 tgaaaaaaga gttgttagcg gttctcgaga tgccactctt agggtttggg atattgagac
     2521 aggccagtgt ttacatgttt tgatgggtca tgttgcagca gtccgctgtg ttcaatatga
     2581 tggcaggagg gttgttagtg gagcatatga ttttatggta aaggtgtggg atccagagac
     2641 tgaaacctgt ctacacacgt tgcaggggca tactaataga gtctattcat tacagtttga
     2701 tggtatccat gtggtgagtg gatctcttga tacatcaatc cgtgtttggg atgtggagac
     2761 agggaattgc attcacacgt taacagggca ccagtcgtta acaagtggaa tggaactcaa
     2821 agacaatatt cttgtctctg ggaatgcaga ttctacagtt aaaatctggg atatcaaaac
     2881 aggacagtgt ttacaaacat tgcaaggtcc caacaagcat cagagtgctg tgacctgttt
     2941 acagttcaac aagaactttg taattaccag ctcagatgat ggaactgtaa aactatggga
     3001 cttgaaaacg ggtgaattta ttcgaaacct agtcacattg gagagtgggg ggagtggggg
     3061 agttgtgtgg cggatcagag cctcaaacac aaagctggtg tgtgcagttg ggagtcggaa
     3121 tgggactgaa gaaaccaagc tgctggtgct ggactttgat gtggacatga agtgaagagc
     3181 agggcccata agggcgaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga
     3241 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3301 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3361 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3421 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3481 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3541 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3601 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3661 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3721 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3781 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3841 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3901 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3961 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4021 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4081 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4141 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4201 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4261 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4321 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4381 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4441 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4501 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4561 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4621 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4681 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4741 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4801 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4861 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4921 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4981 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5041 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5101 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5161 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5221 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5281 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5341 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5401 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5461 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5521 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5581 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5641 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5701 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5761 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5821 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5881 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5941 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6001 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6061 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6121 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6181 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6241 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6301 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6361 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6421 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6481 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6541 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6601 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6661 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6721 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6781 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6841 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6901 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6961 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7021 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7081 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7141 ggccgcagtg
//
