LOCUS       sequence-72795-e        6609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mouse folliculin (FLCN) fused to GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6609)
  AUTHORS   Petit CS, Roczniak-Ferguson A, Ferguson SM
  TITLE     Recruitment of folliculin to lysosomes supports the amino 
            acid-dependent activation of Rag GTPases.
  JOURNAL   J Cell Biol. 2013 Sep 30;202(7):1107-1122.
  PUBMED    24081491
REFERENCE   2  (bases 1 to 6609)
  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"; journalName: "J Cell 
            Biol."; date: "2013-09-30"; volume: "202(7)"; pages: "1107-1122"
COMMENT     Sequence Label: pEGFP-N1-FLCN
FEATURES             Location/Qualifiers
     source          1..6609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2408..2417
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2414..2416,2417..2419,2420..3133)
                     /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(2414..2416,2417..2419,2420..3130)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2459..2480)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2720..2739)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3067..3088
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3256..3377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3293..3312)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3347..3366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3384..3839)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3521..3542)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3733..3752
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3866..3970
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3972..4329
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3972..4163
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3995..4015)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4180..4315
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4364..5158
                     /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             4364..5158
                     /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(4418..4437)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5028..5047
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5346..5365)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5390..5437
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5766..6354
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6255..6274
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcagcacc atgaacgcca tagtcgccct
      661 ctgccacttc tgcgagctcc atggcccccg cacgctcttc tgcacggaag ttctacacgc
      721 tcccctgccc cagggggccg gaagtgggga cagtcctgac caggttgagc aggctgagga
      781 ggaggagggt ggcattcaga tgagcagccg ggtccgtgcc cacagcccag ccgagggtgc
      841 cagcagtgag tccagcagcc cggggcccaa gaagtcggac atgtgtgagg gctgccggtc
      901 acttgccgta gggcacccag gctatatcag tcatgataaa gagacctcta ttaagtacgt
      961 cagtcaccag caccccaacc acccgcagct cttcagcatc gtccgccagg cctgtgtccg
     1021 gagcctgagc tgtgaggtat gccctggtcg tgaaggcccc atcttctttg gtgatgagca
     1081 gcacggcttt gtgttcagcc acaccttctt catcaaagac agcctggcca gaggcttcca
     1141 gcgctggtac agcatcatcg ccatcatgat ggatcggatc tacctcatca actcctggcc
     1201 cttcctgctg gggaggatcc gcggcatcat cagtgagctc caggccaagg ccttcaaggt
     1261 gtttgaggca gagcagtttg gatgtccaca gcgtgcccag aggatgaaca ctgccttcac
     1321 gcccttcctg caccagagga acggcaacgc tgcccgctct ctgacctcct tgaccagtga
     1381 tgacaacttg tgggcgtgtc tgcacacttc ctttgcctgg ctcctgaagg catgcggtag
     1441 caggctgaca gaaaagctct tagagggcgc tcccacagag gacaccctgg tccagatgga
     1501 gaagcttgct gacttggagg aagaatcaga aagttgggac aattctgagg ctgaggagga
     1561 ggagaaagct cctgttacac cagagggtgc tgaagggcga gagctgacca gttgcccaac
     1621 agagtcatcc tttctctcag cctgtgggag ctggcagccc ccaaagctta ccggcttcaa
     1681 gtctcttcga cacatgagac aggtcttggg tgctccatcc ttccgtatgt tggcttggca
     1741 tgtcctcatg gggaatcagg tgatctggaa aagcagagat gtgaacctgg tccattcagc
     1801 gtttgaagtc ctccggacca tgctgcctgt gggctgtgtc cgcatcatcc cttacagcag
     1861 ccagtatgag gaggcctatc gctgcaactt cctggggctc agccctcccg tgcctatccc
     1921 tgcccatgtt ctggcctcag agttcgtagt tgtcgtggag gtccacacgg ccactcgctc
     1981 aaacctccac cctgctgggt gcgaggatga ccagtccctc agcaagtatg agtttgtggt
     2041 gaccagtggt agccctgtgg ctgcagacag agttgggccc actatcctga ataagattga
     2101 agcagctctg accaaccaga acctgtctgt ggatgtggtg gaccaatgtc tcatctgcct
     2161 caaggaggaa tggatgaaca aagtgaaagt cctgtttaaa ttcaccaagg tagacagtcg
     2221 ccccaaggag gacacacaga agctcctaag cgtcctaggc gcatcagagg aggacaacgt
     2281 caaactgctg aagttctgga tgacgggact gagcaaaacc tacaagtccc atctcatgtc
     2341 caccgtccga agccccacag ctacagagtc acggagcacg gtaccgcggg cccgggatcc
     2401 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2461 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2521 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2581 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2641 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2701 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2761 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2821 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2881 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2941 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     3001 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     3061 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     3121 gctgtacaag taaagcggcc gcgactctag atcataatca gccataccac atttgtagag
     3181 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3241 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3301 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3361 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3421 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3481 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3541 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3601 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3661 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3721 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3781 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3841 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3901 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3961 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     4021 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4081 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4141 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4201 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4261 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4321 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4381 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4441 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4501 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4561 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4621 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4681 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4741 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4801 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4861 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4921 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4981 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5041 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5101 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5161 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5221 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5281 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     5341 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5401 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5461 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5521 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5581 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     5641 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5701 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5761 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5821 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5881 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     5941 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6001 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6061 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6121 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6181 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6241 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6301 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6361 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6421 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatcc tggtactcag
     6481 actcactccc ttctactgct ctacctctcc ttctactacc ctaccctcta cctctactaa
     6541 ctactactag ctggagcgct tctactctac tactgaatct cactactact ctcctctaat
     6601 ataactcct
//
