LOCUS       sequence-347651-        6925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pcDNA3-FLAG_FBXL7 (R310H).
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 (R310H)
FEATURES             Location/Qualifiers
     source          1..6925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        130..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     664..684
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     758..777
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        758..776
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             842..865
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2373..2391)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2374..2391)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2411..2428)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2417..2641
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2687..3115
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2774..2793)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2984..3005
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3124..3144)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3129..3458
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3309..3444
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3371..3390
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3525..4319
                     /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(3579..3598)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4189..4208
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4493..4614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4530..4549)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4584..4603
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4663..4679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4663..4679)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4676..4698)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4687..4703
                     /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(4711..4717,4718..4735,4736..4741))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4756..4777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4894..4911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5065..5653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5145..5164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5824..6615,6616..6684))
                     /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     6447..6466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6685..6789)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6864..6883)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
       61 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
      121 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      721 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      781 cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcat
      841 ggactacaag gacgacgatg acaagggcgc gaacaatggc aaacagtacg gcagtgaggg
      901 caaaggcagc tcgagcatct catctgacgt gagttcaagt acagatcaca cgcccactaa
      961 agcccagaag aatgtggcta ccagcgaaga ctccgacctg agcatgcgca cactgagcac
     1021 gcccagccca gccctgatat gtccaccgaa tctcccagga tttcagaatg gaaggggctc
     1081 gtccacctcc tcgtcctcca tcaccgggga gacggtggcc atggtgcact ccccgccccc
     1141 gacccgcctc acacacccgc tcatccggct cgcctccaga ccccagaagg agcaggccag
     1201 catagaccgg ctcccggacc actccatggt gcagatcttc tccttcctgc ccaccaacca
     1261 gctgtgccgc tgcgcgcgag tgtgccgccg ctggtacaac ctggcctggg acccgcggct
     1321 ctggaggact atccgcctga cgggcgagac catcaacgtg gaccgcgccc tcaaggtgct
     1381 gacccgcaga ctctgccagg acacccccaa cgtgtgtctc atgctggaaa ccgtaactgt
     1441 cagtggctgc aggcggctca cagaccgagg gctgtacacc atcgcccagt gctgccccga
     1501 actgaggcga ctggaagtct caggctgtta caatatctcc aacgaggccg tctttgatgt
     1561 ggtgtccctc tgccctaatc tggagcacct ggatgtgtca ggatgctcca aagtgacctg
     1621 catcagcttg acccgggagg cctccattaa actgtcaccc ttgcatggca aacagatttc
     1681 catccgctac ctggacatga cggactgctt cgtgctggag gacgaaggcc tgcacaccat
     1741 cgcggcgcac tgcacgcagc tcacccacct ctacctgcgc cgctgcgtcc acctgaccga
     1801 cgaaggcctg cgctacctgg tgatctactg cgcctccatc aaggagctga gcgtcagcga
     1861 ctgccgcttc gtcagcgact tcggcctgcg ggagatcgcc aagctggagt cccgcctgcg
     1921 gtacctgagc atcgcgcact gcggccgggt caccgacgtg ggcatccgct acgtggccaa
     1981 gtactgcagc aagctgcgct acctcaacgc gaggggctgc gagggcatca cggaccacgg
     2041 tgtggagtac ctcgccaaga actgcaccaa actcaaatcc ctggatatcg gcaaatgccc
     2101 tttggtatcc gacacgggcc tggagtgcct ggccctgaac tgcttcaacc tcaagcggct
     2161 cagcctcaag tcctgcgaga gcatcaccgg ccagggcttg cagatcgtgg ccgccaactg
     2221 ctttgacctc cagacgctga atgtccagga ctgcgaggtc tccgtggagg ccctgcgctt
     2281 tgtcaaacgc cactgcaagc gctgcgtcat cgagcacacc aacccggctt tcttctgagc
     2341 ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct
     2401 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2461 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2521 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2581 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2641 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2701 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2761 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2821 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2881 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2941 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3001 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3061 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3121 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3181 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3241 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3301 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3361 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3421 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3481 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3541 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3601 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3661 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3721 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3781 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3841 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3901 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3961 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4021 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4081 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4141 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4201 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4261 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4321 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4381 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4441 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4501 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4561 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4621 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4681 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4741 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4801 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4861 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4921 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4981 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5041 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5101 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5161 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5221 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5281 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5341 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5401 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5461 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5521 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5581 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5641 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5701 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5761 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5821 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5881 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5941 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6001 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6061 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6121 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6181 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6241 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6301 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6361 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6421 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6481 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6541 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6601 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6661 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6721 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6781 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     6841 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     6901 ctgctccctg cttgtgtgtt ggagg
//
