LOCUS       sequence-345391-        6925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pcDNA3-FLAG_FBXL7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6925)
  AUTHORS   Moro L, Simoneschi D, Kurz E, Arbini AA, Jang S, Guaragnella N, 
            Giannattasio S, Wang W, Chen YA, Pires G, Dang A, Hernandez E, Kapur
            P, Mishra A, Tsirigos A, Miller G, Hsieh JT, Pagano M
  TITLE     Epigenetic silencing of the ubiquitin ligase subunit FBXL7 impairs 
            c-SRC degradation and promotes epithelial-to-mesenchymal transition 
            and metastasis.
  JOURNAL   Nat Cell Biol. 2020 Sep;22(9):1130-1142. doi: 
            10.1038/s41556-020-0560-6. Epub 2020 Aug 24.
  PUBMED    32839549
REFERENCE   2  (bases 1 to 6925)
  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: 
            "10.1038/s41556-020-0560-6"; journalName: "Nat Cell Biol"; date: 
            "2020-09"; volume: "22"; issue: "9"; pages: "1130-1142"
COMMENT     Sequence Label: pcDNA3-FLAG_FBXL7
FEATURES             Location/Qualifiers
     source          1..6925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        156..535
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        536..739
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     690..710
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     784..803
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        784..802
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             868..891
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2399..2417)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2400..2417)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2437..2454)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2443..2667
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2713..3141
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2800..2819)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3010..3031
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3150..3170)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3155..3484
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3335..3470
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3397..3416
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3551..4345
                     /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(3605..3624)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4215..4234
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4519..4640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4556..4575)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4610..4629
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4689..4705)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4689..4705)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4702..4724)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4713..4729
                     /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(4737..4743,4744..4761,4762..4767))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4782..4803
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4920..4937)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5091..5679)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5171..5190)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5850..6641,6642..6710))
                     /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     6473..6492
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6711..6815)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6890..6909)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
       61 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
      121 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
      181 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      241 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      301 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      361 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      421 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      481 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      541 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      601 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      661 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      721 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
      781 aattaatacg actcactata gggagaccca agcttggtac cgagctcgga tccactagta
      841 acggccgcca gtgtgctgga attcatggac tacaaggacg acgatgacaa gggcgcgaac
      901 aatggcaaac agtacggcag tgagggcaaa ggcagctcga gcatctcatc tgacgtgagt
      961 tcaagtacag atcacacgcc cactaaagcc cagaagaatg tggctaccag cgaagactcc
     1021 gacctgagca tgcgcacact gagcacgccc agcccagccc tgatatgtcc accgaatctc
     1081 ccaggatttc agaatggaag gggctcgtcc acctcctcgt cctccatcac cggggagacg
     1141 gtggccatgg tgcactcccc gcccccgacc cgcctcacac acccgctcat ccggctcgcc
     1201 tccagacccc agaaggagca ggccagcata gaccggctcc cggaccactc catggtgcag
     1261 atcttctcct tcctgcccac caaccagctg tgccgctgcg cgcgagtgtg ccgccgctgg
     1321 tacaacctgg cctgggaccc gcggctctgg aggactatcc gcctgacggg cgagaccatc
     1381 aacgtggacc gcgccctcaa ggtgctgacc cgcagactct gccaggacac ccccaacgtg
     1441 tgtctcatgc tggaaaccgt aactgtcagt ggctgcaggc ggctcacaga ccgagggctg
     1501 tacaccatcg cccagtgctg ccccgaactg aggcgactgg aagtctcagg ctgttacaat
     1561 atctccaacg aggccgtctt tgatgtggtg tccctctgcc ctaatctgga gcacctggat
     1621 gtgtcaggat gctccaaagt gacctgcatc agcttgaccc gggaggcctc cattaaactg
     1681 tcacccttgc atggcaaaca gatttccatc cgctacctgg acatgacgga ctgcttcgtg
     1741 ctggaggacg aaggcctgca caccatcgcg gcgcactgca cgcagctcac ccacctctac
     1801 ctgcgccgct gcgtccgcct gaccgacgaa ggcctgcgct acctggtgat ctactgcgcc
     1861 tccatcaagg agctgagcgt cagcgactgc cgcttcgtca gcgacttcgg cctgcgggag
     1921 atcgccaagc tggagtcccg cctgcggtac ctgagcatcg cgcactgcgg ccgggtcacc
     1981 gacgtgggca tccgctacgt ggccaagtac tgcagcaagc tgcgctacct caacgcgagg
     2041 ggctgcgagg gcatcacgga ccacggtgtg gagtacctcg ccaagaactg caccaaactc
     2101 aaatccctgg atatcggcaa atgccctttg gtatccgaca cgggcctgga gtgcctggcc
     2161 ctgaactgct tcaacctcaa gcggctcagc ctcaagtcct gcgagagcat caccggccag
     2221 ggcttgcaga tcgtggccgc caactgcttt gacctccaga cgctgaatgt ccaggactgc
     2281 gaggtctccg tggaggccct gcgctttgtc aaacgccact gcaagcgctg cgtcatcgag
     2341 cacaccaacc cggctttctt ctgagcggcc gctcgagcat gcatctagag ggccctattc
     2401 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     2461 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2521 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2581 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2641 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     2701 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2761 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2821 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2881 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2941 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3001 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3061 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3121 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3181 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3241 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3301 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3361 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3421 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3481 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     3541 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3601 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3661 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3721 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3781 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3841 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3901 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3961 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4021 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4081 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4141 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4201 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4261 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4321 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4381 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4441 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4501 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4561 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4621 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4681 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4741 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4801 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgtat
     4861 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4921 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4981 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5041 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5101 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5161 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5221 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5281 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5341 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5401 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5461 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5521 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5581 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5641 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5701 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5761 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5821 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5881 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5941 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6001 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6061 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6121 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6181 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6241 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6301 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6361 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6421 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6481 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6541 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6601 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6661 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6721 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6781 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6841 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     6901 ctgatgccgc atagttaagc cagta
//
