LOCUS       sequence-60643-e        6062 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 6062)
  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 6062)
  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-delta30-TFEB
FEATURES             Location/Qualifiers
     source          1..6062
                     /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      628..637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2002..2011
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2008..2010,2011..2013,2014..2727)
                     /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(2008..2010,2011..2013,2014..2724)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2053..2074)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2314..2333)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2661..2682
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2850..2971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2887..2906)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2941..2960
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2978..3433)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3115..3136)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3327..3346
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3460..3564
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3566..3923
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3566..3757
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3589..3609)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3774..3909
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3958..4752
                     /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             3958..4752
                     /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(4012..4031)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4622..4641
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4940..4959)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4984..5031
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5360..5948
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5849..5868
                     /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 caagcttgcc accatggatt acatgcagca gcagcagcag
      661 cagcaacagc agcagctcgg agggccgccc accccggcca tcaatacccc cgtccacttc
      721 cagtcgccac cacctgtgcc tggggaggtg ttgaaggtgc agtcctacct ggagaatccc
      781 acatcctacc atctgcagca gtcgcagcat cagaaggtgc gggagtacct gtccgagacc
      841 tatgggaaca agtttgctgc ccacatcagc ccagcccagg gctctccgaa acccccacca
      901 gccgcctccc caggggtgcg agctggacac gtgctgtcct cctccgctgg caacagtgct
      961 cccaatagcc ccatggccat gctgcacatt ggctccaacc ctgagaggga gttggatgat
     1021 gtcattgaca acattatgcg tctggacgat gtccttggct acatcaatcc tgaaatgcag
     1081 atgcccaaca cgctacccct gtccagcagc cacctgaatg tgtacagcag cgacccccag
     1141 gtcacagcct ccctggtggg cgtcaccagc agctcctgcc ctgcggacct gacccagaag
     1201 cgagagctca cagatgctga gagcagggcc ctggccaagg agcggcagaa gaaagacaat
     1261 cacaacttaa ttgaaaggag acgaaggttc aacatcaatg accgcatcaa ggagttggga
     1321 atgctgatcc ccaaggccaa tgacctggac gtgcgctgga acaagggcac catcctcaag
     1381 gcctctgtgg attacatccg gaggatgcag aaggacctgc aaaagtccag ggagctggag
     1441 aaccactctc gccgcctgga gatgaccaac aagcagctct ggctccgtat ccaggagctg
     1501 gagatgcagg ctcgagtgca cggcctccct accacctccc cgtccggcat gaacatggct
     1561 gagctggccc agcaggtggt gaagcaggag ctgcctagcg aagagggccc aggggaggcc
     1621 ctgatgctgg gggctgaggt ccctgaccct gagccactgc cagctctgcc cccgcaagcc
     1681 ccgctgcccc tgcccaccca gccaccatcc ccattccatc acctggactt cagccacagc
     1741 ctgagctttg ggggcaggga ggacgagggt cccccgggct accccgaacc cctggcgccg
     1801 gggcatggct ccccattccc cagcctgtcc aagaaggatc tggacctcat gctcctggac
     1861 gactcactgc taccgctggc ctctgatcca cttctgtcca ccatgtcccc cgaggcctcc
     1921 aaggccagca gccgccggag cagcttcagc atggaggagg gcgatgtgct gaaggtaccg
     1981 cgggcccggg atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     2041 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     2101 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     2161 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     2221 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     2281 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     2341 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     2401 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     2461 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     2521 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     2581 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     2641 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2701 ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata
     2761 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2821 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2881 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2941 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     3001 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3061 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3121 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3181 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3241 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3301 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3361 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3421 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3481 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3541 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3601 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3661 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3721 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3781 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3841 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3901 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     3961 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4021 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4081 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4141 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4201 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4261 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4321 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4381 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4441 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4501 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4561 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4621 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4681 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4741 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4801 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4861 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4921 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4981 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     5041 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     5101 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     5161 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     5221 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5281 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5341 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5401 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5461 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     5521 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5581 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5641 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5701 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5761 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5821 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5881 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5941 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6001 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     6061 at
//
