LOCUS       sequence-473146-        7599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of FBXW7alpha in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7599)
  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 7599)
  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 FBXW7alpha
FEATURES             Location/Qualifiers
     source          1..7599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(166..270)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(345..364)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        536..915
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        916..1119
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1070..1090
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1164..1183
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1164..1182
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1224..1247
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3491..3508)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3497..3721
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3767..4195
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3854..3873)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4064..4085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4204..4224)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4209..4538
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4389..4524
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4451..4470
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4605..5399
                     /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(4659..4678)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5269..5288
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5573..5694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5610..5629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5664..5683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5743..5759)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5743..5759)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5756..5778)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5767..5783
                     /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(5791..5797,5798..5815,5816..5821))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5836..5857
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5974..5991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6145..6733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6225..6244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6904..7599,1..96,97..165))
                     /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     7527..7546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
       61 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
      121 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
      181 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
      241 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
      301 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
      361 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      421 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      481 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      541 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      601 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      661 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      721 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      781 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      841 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      901 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      961 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     1021 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     1081 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
     1141 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
     1201 cgtttaaact taagcttacc atggactaca aggacgacga tgacaaggta ccaggatcca
     1261 gtgtggtgga attcgccctt cagaagcttt tggaaatgaa tcaggaactg ctctctgtgg
     1321 gcagcaaaag acgacgaact ggaggctctc tgagaggtaa cccttcctca agccaggtag
     1381 atgaagaaca gatgaatcgt gtggtagagg aggagcagca acagcaactc agacaacaag
     1441 aggaggagca cactgcaagg aatggtgaag ttgttggagt agaacctaga cctggaggcc
     1501 aaaatgattc ccagcaagga cagttggaag aaaacaataa tagatttatt tcggtagatg
     1561 aggactcctc aggaaaccaa gaagaacaag aggaagatga agaacatgct ggtgaacaag
     1621 atgaggagga tgaggaggag gaggagatgg accaggagag tgacgatttt gatcagtctg
     1681 atgatagtag cagagaagat gaacatacac atactaacag tgtcacgaac accagtagta
     1741 ttgtggacct gcccgttcac caactctcct ccccattcta tacaaaaaca acaaaaatga
     1801 aaagaaagtt ggaccatggt tctgaggtcc gctctttttc tttgggaaag aaaccatgca
     1861 aagtctcaga atatacaagt accactgggc ttgtaccatg ttcagcaaca ccaacaactt
     1921 ttggggacct cagagcagcc aatggccaag ggcaacaacg acgccgaatt acatctgtcc
     1981 agccacctac aggcctccag gaatggctaa aaatgtttca gagctggagt ggaccagaga
     2041 aattgcttgc tttagatgaa ctcattgata gttgtgaacc aacacaagta aaacatatga
     2101 tgcaagtgat agaaccccag tttcaacgag acttcatttc attgctccct aaagagttgg
     2161 cactctatgt gctttcattc ctggaaccca aagacctgct acaagcagct cagacatgtc
     2221 gctactggag aattttggct gaagacaacc ttctctggag agagaaatgc aaagaagagg
     2281 ggattgatga accattgcac atcaagagaa gaaaagtaat aaaaccaggt ttcatacaca
     2341 gtccatggaa aagtgcatac atcagacagc acagaattga tactaactgg aggcgaggag
     2401 aactcaaatc tcctaaggtg ctgaaaggac atgatgatca tgtgatcaca tgcttacagt
     2461 tttgtggtaa ccgaatagtt agtggttctg atgacaacac tttaaaagtt tggtcagcag
     2521 tcacaggcaa atgtctgaga acattagtgg gacatacagg tggagtatgg tcatcacaaa
     2581 tgagagacaa catcatcatt agtggatcta tagatcggac actcaaagtg tggaatgcag
     2641 agactggaga atgtatacac accttatatg ggcatacttc cactgtgcgt tgtatgcatc
     2701 ttcatgaaaa aagagttgtt agcggttctc gagatgccac tcttagggtt tgggatattg
     2761 agacaggcca gtgtttacat gttttgatgg gtcatgttgc agcagtccgc tgtgttcaat
     2821 atgatggcag gagggttgtt agtggagcat atgattttat ggtaaaggtg tgggatccag
     2881 agactgaaac ctgtctacac acgttgcagg ggcatactaa tagagtctat tcattacagt
     2941 ttgatggtat ccatgtggtg agtggatctc ttgatacatc aatccgtgtt tgggatgtgg
     3001 agacagggaa ttgcattcac acgttaacag ggcaccagtc gttaacaagt ggaatggaac
     3061 tcaaagacaa tattcttgtc tctgggaatg cagattctac agttaaaatc tgggatatca
     3121 aaacaggaca gtgtttacaa acattgcaag gtcccaacaa gcatcagagt gctgtgacct
     3181 gtctacagtt caacaagaac tttgtaatta ccagctcaga tgatggaact gtaaaactat
     3241 gggacttgaa aacgggtgaa tttattcgaa acctagtcac attggagagt ggggggagtg
     3301 ggggagttgt gtggcggatc agagcctcaa acacaaagct ggtgtgtgca gttgggagtc
     3361 ggaatgggac tgaagaaacc aagctgctgg tgctggactt tgatgtggac atgaagtgaa
     3421 gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc
     3481 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3541 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3601 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3661 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3721 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3781 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3841 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3901 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3961 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4021 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4081 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4141 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4201 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4261 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4321 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4381 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4441 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4501 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4561 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4621 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4681 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4741 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4801 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4861 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4921 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4981 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5041 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5101 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5161 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5221 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5281 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5341 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5401 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5461 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5521 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5581 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5641 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5701 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5761 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5821 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5881 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5941 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6001 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6061 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6121 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6181 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6241 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6301 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6361 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6421 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6481 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6541 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6601 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6661 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6721 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6781 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6841 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6901 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6961 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7021 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7081 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7141 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7201 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7261 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7321 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7381 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7441 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7501 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7561 gctcatcatt ggaaaacgtt cttcggggcg aaaactctc
//
