LOCUS       sequence-201237-        6449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6449)
  AUTHORS   Roczniak-Ferguson A, Petit CS, Froehlich F, Qian S, Ky J, Angarola 
            B, Walther TC, Ferguson SM
  TITLE     The Transcription Factor TFEB Links mTORC1 Signaling to 
            Transcriptional Control of Lysosome Homeostasis.
  JOURNAL   Sci Signal. 2012 Jun 12;5(228):ra42.
  PUBMED    22692423
REFERENCE   2  (bases 1 to 6449)
  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: "Sci 
            Signal."; date: "2012-06-12"; volume: "5(228)"; pages: "ra42"
COMMENT     Sequence Label: pEGFP-N1-TFE3
FEATURES             Location/Qualifiers
     source          1..6449
                     /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      2329..2338
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2335..2337,2338..2340,2341..3054)
                     /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(2335..2337,2338..2340,2341..3051)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2380..2401)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2641..2660)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2988..3009
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3177..3298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3214..3233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3268..3287
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3305..3760)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3442..3463)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3654..3673
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3787..3891
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3893..4250
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3893..4084
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3916..3936)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4101..4236
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4285..5079
                     /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             4285..5079
                     /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(4339..4358)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4949..4968
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5267..5286)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5311..5358
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5687..6275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6176..6195
                     /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 caagcttgac accatgtctc atgcggccga accagctcgg
      601 gatggcgtag aggccagcgc ggagggccct cgagccgtgt tcgtgctgtt ggaggagcgc
      661 aggccggccg actcggctca gctgctcagc ctgaactctt tgcttccgga atccgggatt
      721 gttgctgaca tagaattaga aaacgtcctt gatcctgaca gcttctacga gctcaaaagc
      781 caacccttac cccttcgctc aagcctccca atatcactgc aggccacacc agccacccca
      841 gctacactct ctgcatcgtc ttctgcaggg ggctccagga cccctgccat gtcgtcatct
      901 tcttcatcga gggtcttgct gcggcagcag ctaatgcggg cccaggcgca ggagcaggag
      961 aggcgtgagc gtcgggaaca ggccgccgcg gctcccttcc ccagtcctgc acctgcctct
     1021 cctgccatct ctgtggttgg cgtctctgct gggggccaca cattgagccg tccaccccct
     1081 gctcaggtgc ccagggaggt gctcaaggtg cagacccatc tggagaaccc aacgcgctac
     1141 cacctgcagc aggcgcgccg gcagcaggtg aaacagtacc tgtccaccac actcgggccc
     1201 aagctggctt cccaggccct caccccaccg ccggggcccg caagtgccca gccactgcct
     1261 gcccctgagg ctgcccacac taccggcccc acaggcagtg cgcccaacag ccccatggcg
     1321 ctgctcacca tcgggtccag ctcagagaag gagattgatg atgtcattga tgagatcatc
     1381 agcctggagt ccagttacaa tgatgaaatg ctcagctatc tgcccggagg caccacagga
     1441 ctgcagctcc ccagcacgct gcctgtgtca gggaatctgc ttgatgtgta cagtagtcaa
     1501 ggcgtggcca caccagccat cactgtcagc aactcctgcc cagctgagct gcccaacatc
     1561 aaacgggaga tctctgagac cgaggcaaag gcccttttga aggaacggca gaagaaagac
     1621 aatcacaacc taattgagcg tcgcaggcga ttcaacatta acgacaggat caaggaactg
     1681 ggcactctca tccctaagtc cagtgacccg gagatgcgct ggaacaaggg caccatcctg
     1741 aaggcctctg tggattatat ccgcaagctg cagaaggagc agcagcgctc caaagacctg
     1801 gagagccggc agcgatccct ggagcaggcc aaccgcagcc tgcagctccg aattcaggaa
     1861 ctagaactgc aggcccagat ccatggcctg ccagtgcctc ccactccagg gctgctttcc
     1921 ttggccacga cttcggcttc tgacagcctc aagccagagc agctggacat tgaggaggag
     1981 ggcaggccag gcgcagcaac gttccatgta ggggggggac ctgcccagaa tgctccccat
     2041 cagcagcccc ctgcaccgcc ctcagatgcc cttctggacc tgcactttcc cagcgaccac
     2101 ctgggggacc tgggagaccc cttccacctg gggctggagg acattctgat ggaggaggag
     2161 gagggggtgg tgggaggact gtcggggggt gccctgtccc cactgcgggc tgcctccgat
     2221 cccctgctct cttcagtgtc ccctgctgtc tccaaggcca gcagccgccg cagcagcttc
     2281 agcatggaag aggagtccac ggtaccgcgg gcccgggatc caccggtcgc caccatggtg
     2341 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2401 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2461 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2521 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2581 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2641 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2701 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2761 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2821 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2881 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2941 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3001 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     3061 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     3121 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     3181 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3241 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     3301 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3361 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3421 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3481 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3541 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3601 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3661 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3721 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     3781 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3841 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     3901 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3961 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4021 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4081 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4141 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4201 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     4261 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4321 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4381 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4441 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4501 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4561 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4621 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4681 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4741 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4801 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4861 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4921 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4981 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5041 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5101 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5161 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5221 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     5281 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     5341 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     5401 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     5461 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     5521 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5581 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5641 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5701 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5761 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5821 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5881 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5941 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6001 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6061 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6121 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6181 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6241 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6301 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6361 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     6421 ttagttcata gcccatatat ggagttccg
//
