LOCUS       sequence-345823-        6350 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pEGFP-N1_c-SRC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6350)
  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 6350)
  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: pEGFP-N1_c-SRC
FEATURES             Location/Qualifiers
     source          1..6350
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        93..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..600
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     551..571
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2322..2331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2328..2330,2331..2333,2334..3047)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2328..2330,2331..2333,2334..3044)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2373..2394)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2634..2653)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2981..3002
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3170..3291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3207..3226)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3261..3280
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3298..3753)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3435..3456)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3647..3666
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3780..3884
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3886..4243
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3886..4077
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3909..3929)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4094..4229
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4278..5072
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4278..5072
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4332..4351)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4942..4961
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5260..5279)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5304..5351
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5680..6268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6169..6188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
       61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      601 ggtttagtga accgtcagat ccgctagcgc taccggactc agatctcgag atgggtagca
      661 acaagagcaa gcccaaggat gccagccagc ggcgccgcag cctggagccc gccgagaacg
      721 tgcacggcgc tggcgggggc gctttccccg cctcgcagac ccccagcaag ccagcctcgg
      781 ccgacggcca ccgcggcccc agcgcggcct tcgcccccgc ggccgccgag cccaagctgt
      841 tcggaggctt caactcctcg gacaccgtca cctccccgca gagggcgggc ccgctggccg
      901 gtggagtgac cacctttgtg gccctctatg actatgagtc taggacggag acagacctgt
      961 ccttcaagaa aggcgagcgg ctccagattg tcaacaacac agagggagac tggtggctgg
     1021 cccactcgct cagcacagga cagacaggct acatccccag caactacgtg gcgccctccg
     1081 actccatcca ggctgaggag tggtattttg gcaagatcac cagacgggag tcagagcggt
     1141 tactgctcaa tgcagagaac ccgagaggga ccttcctcgt gcgagaaagt gagaccacga
     1201 aaggtgccta ctgcctctca gtgtctgact tcgacaacgc caagggcctc aacgtgaagc
     1261 actacaagat ccgcaagctg gacagcggcg gcttctacat cacctcccgc acccagttca
     1321 acagcctgca gcagctggtg gcctactact ccaaacacgc cgatggcctg tgccaccgcc
     1381 tcaccaccgt gtgccccacg tccaagccgc agactcaggg cctggccaag gatgcctggg
     1441 agatccctcg ggagtcgctg cggctggagg tcaagctggg ccagggctgc tttggcgagg
     1501 tgtggatggg gacctggaac ggtaccacca gggtggccat caaaaccctg aagcctggca
     1561 cgatgtctcc agaggccttc ctgcaggagg cccaggtcat gaagaagctg aggcatgaga
     1621 agctggtgca gttgtatgct gtggtttcag aggagcccat ttacatcgtc acggagtaca
     1681 tgagcaaggg gagtttgctg gactttctca agggggagac aggcaagtac ctgcggctgc
     1741 ctcagctggt ggacatggct gctcagatcg cctcaggcat ggcgtacgtg gagcggatga
     1801 actacgtcca ccgggacctt cgtgcagcca acatcctggt gggagagaac ctggtgtgca
     1861 aagtggccga ctttgggctg gctcggctca ttgaagacaa tgagtacacg gcgcggcaag
     1921 gtgccaaatt ccccatcaag tggacggctc cagaagctgc cctctatggc cgcttcacca
     1981 tcaagtcgga cgtgtggtcc ttcgggatcc tgctgactga gctcaccaca aagggacggg
     2041 tgccctaccc tgggatggtg aaccgcgagg tgctggacca ggtggagcgg ggctaccgga
     2101 tgccctgccc gccggagtgt cccgagtccc tgcacgacct catgtgccag tgctggcgga
     2161 aggagcctga ggagcggccc accttcgagt acctgcaggc cttcctggag gactacttca
     2221 cgtccaccga gccccagtac cagcccgggg agaacctcgg atcaggatca aagcttcgaa
     2281 ttctgcagtc gacggtaccg cgggcccggg atccaccggt cgccaccatg gtgagcaagg
     2341 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2401 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     2461 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2521 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     2581 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     2641 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2701 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2761 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     2821 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     2881 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     2941 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     3001 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact
     3061 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3121 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3181 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3241 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     3301 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3361 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     3421 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     3481 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     3541 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     3601 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3661 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3721 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     3781 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3841 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     3901 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3961 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4021 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4081 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4141 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4201 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     4261 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4321 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4381 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4441 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4501 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4561 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4621 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4681 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4741 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4801 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4861 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4921 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4981 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5041 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5101 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5161 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5221 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     5281 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     5341 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     5401 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     5461 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     5521 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5581 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5641 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5701 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5761 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5821 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5881 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5941 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6001 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6061 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6121 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6181 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6241 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6301 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
//
