LOCUS       sequence-307896-        6453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human FLCN tagged with eGFP at C terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6453)
  AUTHORS   Tsun ZY, Bar-Peled L, Chantranupong L, Zoncu R, Wang T, Kim C, 
            Spooner E, Sabatini DM
  TITLE     The folliculin tumor suppressor is a GAP for the RagC/D GTPases that
            signal amino acid levels to mTORC1.
  JOURNAL   Mol Cell. 2013 Nov 21;52(4):495-505. doi: 
            10.1016/j.molcel.2013.09.016. Epub 2013 Oct 3.
  PUBMED    24095279
REFERENCE   2  (bases 1 to 6453)
  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: 
            "10.1016/j.molcel.2013.09.016"; journalName: "Mol Cell"; date: 
            "2013-11-21- 21"; volume: "52"; issue: "4"; pages: "495-505"
COMMENT     Sequence Label: FLCN-eGFP
FEATURES             Location/Qualifiers
     source          1..6453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2333..2342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2339..2341,2342..2344,2345..3058)
                     /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(2339..2341,2342..2344,2345..3055)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2384..2405)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2645..2664)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2992..3013
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3181..3302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3218..3237)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3272..3291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3309..3764)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3446..3467)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3658..3677
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3791..3895
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3897..4254
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3897..4088
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3920..3940)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4105..4240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4289..5083
                     /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             4289..5083
                     /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(4343..4362)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4953..4972
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5271..5290)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5315..5362
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5691..6279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6180..6199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcgccacc atgaatgcca tcgtggctct
      601 ctgccacttc tgcgagctcc acggcccccg cactctcttc tgcacggagg tgctgcacgc
      661 cccacttcct caaggggatg ggaatgagga cagtcccggc cagggtgagc aggcggaaga
      721 agaggaaggt ggcattcaga tgaacagtcg gatgcgtgcg cacagccccg cagagggggc
      781 cagcgtcgag tccagcagcc cggggcccaa aaagtcggac atgtgcgagg gctgccggtc
      841 acttgctgca gggcacccgg gatatatcag ccatgataaa gagacctcca ttaaatacgt
      901 cagccaccag caccccagcc acccccagct cttcagcatt gtccgccagg cctgtgtccg
      961 gagcctgagc tgtgaggtct gccctggccg tgaaggcccc atcttcttcg gagatgagca
     1021 gcacggcttt gtgttcagcc acaccttctt catcaaggac agcctggcca ggggcttcca
     1081 gcgctggtac agcatcatca ccatcatgat ggaccggatc tacctcatca actcctggcc
     1141 cttcctgctg gggaaggtcc ggggaatcat cgatgagctc cagggcaagg cgctcaaggt
     1201 gtttgaggca gagcagtttg gatgcccaca gcgtgctcag aggatgaaca cagccttcac
     1261 gccattccta caccagagga acggcaacgc cgcccgctcg ctgacatcgc tgacaagtga
     1321 tgacaacctg tgggcgtgcc tgcacacctc ctttgcctgg ctcctgaagg cgtgtggcag
     1381 ccggctgacc gagaagctcc tggaaggtgc tccgaccgag gataccttgg tccagatgga
     1441 gaagctcgct gatttagaag aggaatcaga aagctgggac aactctgagg ctgaagagga
     1501 ggagaaagcc cctgtgttgc cagagagtac agaagggcgg gagctgaccc agggcccggc
     1561 agagtcctcc tctctctcag gctgtgggag ctggcagccc cggaagctgc cagtcttcaa
     1621 gtccctccgg cacatgaggc aggtcctggg tgccccttct ttccgcatgc tggcctggca
     1681 cgttctcatg gggaaccagg tgatctggaa aagcagagac gtggacctcg tccagtcagc
     1741 ttttgaagta cttcggacca tgcttcccgt gggctgcgtc cgcatcatcc catacagcag
     1801 ccagtacgag gaggcctatc ggtgcaactt cctggggctc agcccgcacg tgcagatccc
     1861 cccccacgtg ctctcctcag agtttgctgt catcgtggag gtccacgcag ccgcacgttc
     1921 caccctccac cctgtggggt gtgaggatga ccagtctctc agcaagtacg agtttgtggt
     1981 gaccagtggg agccctgtag ctgcagaccg agtgggcccc accatcctga ataagattga
     2041 agcggctctg accaaccaga acctgtctgt ggatgtggtg gaccagtgcc tcgtctgcct
     2101 caaggaggag tggatgaaca aagtgaaggt gctttttaag ttcaccaagg tggacagtcg
     2161 ccccaaagag gacacacaga agctgctgag catcctgggt gcgtccgagg aggacaatgt
     2221 caagctgctg aagttctgga tgactggcct gagcaagacc tacaagtcac acctcatgtc
     2281 cacggtccgc agccccacag cctcggagtc tcggaaccgg gatccaccgg tcgccaccat
     2341 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2401 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     2461 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2521 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     2581 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2641 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2701 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2761 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2821 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2881 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2941 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3001 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     3061 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3121 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3181 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3241 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3301 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3361 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3421 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3481 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3541 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3601 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3661 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3721 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3781 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3841 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3901 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3961 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4081 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4141 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4201 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     4261 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4321 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4381 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4441 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4501 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4561 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4621 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4681 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4741 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4801 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4861 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4921 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4981 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5041 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5101 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5161 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5221 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     5281 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5341 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5401 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5461 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5521 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5581 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5641 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5701 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5761 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5821 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5881 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5941 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6001 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6061 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6121 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6181 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6241 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6301 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6361 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
     6421 gtcattagtt catagcccat atatggagtt ccg
//
