LOCUS       sequence-72800-e        7178 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of mouse folliculin (FLCN) fused to tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7178)
  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 7178)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: ptdTomato-N1-FLCN
FEATURES             Location/Qualifiers
     source          1..7178
                     /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             2414..3844
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     polyA_signal    3966..4087
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4003..4022)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4057..4076
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4094..4549)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4231..4252)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4443..4462
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4576..4680
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4682..5039
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4682..4873
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4705..4725)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4890..5025
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5074..5868
                     /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             5074..5868
                     /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(5128..5147)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5738..5757
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6056..6075)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6100..6147
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6476..7064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6965..6984
                     /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 ggaggtcatc aaagagttca tgcgcttcaa
     2461 ggtgcgcatg gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     2521 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc
     2581 cttcgcctgg gacatcctgt ccccccagtt catgtacggc tccaaggcgt acgtgaagca
     2641 ccccgccgac atccccgatt acaagaagct gtccttcccc gagggcttca agtgggagcg
     2701 cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc caggactcct ccctgcagga
     2761 cggcacgctg atctacaagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt
     2821 aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg
     2881 cgtgctgaag ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt
     2941 ggagttcaag accatctaca tggccaagaa gcccgtgcaa ctgcccggct actactacgt
     3001 ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     3061 gcgctccgag ggccgccacc acctgttcct ggggcatggc accggcagca ccggcagcgg
     3121 cagctccggc accgcctcct ccgaggacaa caacatggcc gtcatcaaag agttcatgcg
     3181 cttcaaggtg cgcatggagg gctccatgaa cggccacgag ttcgagatcg agggcgaggg
     3241 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc
     3301 cctgcccttc gcctgggaca tcctgtcccc ccagttcatg tacggctcca aggcgtacgt
     3361 gaagcacccc gccgacatcc ccgattacaa gaagctgtcc ttccccgagg gcttcaagtg
     3421 ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc gtgacccagg actcctccct
     3481 gcaggacggc acgctgatct acaaggtgaa gatgcgcggc accaacttcc cccccgacgg
     3541 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc tgtacccccg
     3601 cgacggcgtg ctgaagggcg agatccacca ggccctgaag ctgaaggacg gcggccacta
     3661 cctggtggag ttcaagacca tctacatggc caagaagccc gtgcaactgc ccggctacta
     3721 ctacgtggac accaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     3781 gtacgagcgc tccgagggcc gccaccacct gttcctgtac ggcatggacg agctgtacaa
     3841 gtaggcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     3901 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3961 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4021 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4081 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     4141 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4201 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4261 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     4321 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     4381 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4441 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4501 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     4561 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4621 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4681 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4741 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4801 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4861 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4921 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4981 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     5041 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5101 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5161 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5221 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     5281 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5341 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5401 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5461 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5521 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5581 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     5641 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5701 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5761 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5821 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5881 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5941 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6001 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     6061 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     6121 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     6181 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     6241 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     6301 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     6361 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6421 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6481 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6541 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6601 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6661 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6721 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6781 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6841 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6901 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6961 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7021 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7081 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     7141 cccctgattc tgtggataac cgtattaccg ccatgcat
//
