LOCUS       sequence-273134-        8257 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TFEB-sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8257)
  AUTHORS   Ishii S, Matsuura A, Itakura E
  TITLE     Identification of a factor controlling lysosomal homeostasis using a
            novel lysosomal trafficking probe.
  JOURNAL   Sci Rep. 2019 Aug 12;9(1):11635. doi: 10.1038/s41598-019-48131-2.
  PUBMED    31406169
REFERENCE   2  (bases 1 to 8257)
  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 Rep."; 
            date: "2019-08-12"; volume: "9(1):11635. doi"; pages: " 
            10.1038/s41598-019-48131-2"
COMMENT     Sequence Label: pMRX-IP TFEB-sfGFP
FEATURES             Location/Qualifiers
     source          1..8257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    128..485
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             550..966
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     896..918
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     934..950
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    976..1350
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1288..1304
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(2813..2815,2816..2818,2819..3532)
                     /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(2813..2815,2816..2818,2819..3529)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2858..2879)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3119..3138)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3466..3487
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(3563..4136,4137..4139,4140..4148)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3729..3746)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3956..3975
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4149..4748
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4149..4168)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4645..4665
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4972..5441
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5571..5588)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5742..6330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5822..5841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6494..7285,7286..7354))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7117..7136
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7355..7459)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7527..7545
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        7647..8026
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt
       61 tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg acccaccacc
      121 gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg tctatgactg
      181 attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg cggacccgtg
      241 gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc agggacttcg
      301 ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg actctttggt
      361 gcacccccct tagaggaggg atatgtggtt ctggtaggag acgagaacct aaaacagttc
      421 ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc gcgtcttgtc
      481 tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat
      541 atgggcccgg gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat
      601 gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc
      661 tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga
      721 gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac
      781 caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag
      841 ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt
      901 gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta
      961 ggcgccccca tatggccata tgagatctta tatggggcac ccccgcccct tgtaaacttc
     1021 cctgaccctg acatgacaag agttactaac agcccctctc tccaagctca cttacaggct
     1081 ctctacttag tccagcacga agtctggaga cctctggcgg cagcctacca agaacaactg
     1141 gaccgaccgg tggtacctca cccttaccga gtcggcgaca cagtgtgggt ccgccgacac
     1201 cagactaaga acctagaacc tcgctggaaa ggaccttaca cagtcctgct gaccaccccc
     1261 accgccctca aagtagacgg catcgcagct tggatacacg ccgcccacgt gaaggctgcc
     1321 gaccccgggg gtggaccatc ctctagactg ccggatcatc gatgaattca ccatggcgtc
     1381 acgcataggg ttgcgcatgc agctcatgcg ggagcaggcg cagcaggagg agcagcggga
     1441 gcgcatgcag caacaggctg tcatgcatta catgcagcag cagcagcagc agcaacagca
     1501 gcagctcgga gggccgccca ccccggccat caataccccc gtccacttcc agtcgccacc
     1561 acctgtgcct ggggaggtgt tgaaggtgca gtcctacctg gagaatccca catcctacca
     1621 tctgcagcag tcgcagcatc agaaggtgcg ggagtacctg tccgagacct atgggaacaa
     1681 gtttgctgcc cacatcagcc cagcccaggg ctctccgaaa cccccaccag ccgcctcccc
     1741 aggggtgcga gctggacacg tgctgtcctc ctccgctggc aacagtgctc ccaatagccc
     1801 catggccatg ctgcacattg gctccaaccc tgagagggag ttggatgatg tcattgacaa
     1861 cattatgcgt ctggacgatg tccttggcta catcaatcct gaaatgcaga tgcccaacac
     1921 gctacccctg tccagcagcc acctgaatgt gtacagcagc gacccccagg tcacagcctc
     1981 cctggtgggc gtcaccagca gctcctgccc tgcggacctg acccagaagc gagagctcac
     2041 agatgctgag agcagggccc tggccaagga gcggcagaag aaagacaatc acaacttaat
     2101 tgaaaggaga cgaaggttca acatcaatga ccgcatcaag gagttgggaa tgctgatccc
     2161 caaggccaat gacctggacg tgcgctggaa caagggcacc atcctcaagg cctctgtgga
     2221 ttacatccgg aggatgcaga aggacctgca aaagtccagg gagctggaga accactctcg
     2281 ccgcctggag atgaccaaca agcagctctg gctccgtatc caggagctgg agatgcaggc
     2341 tcgagtgcac ggcctcccta ccacctcccc gtccggcatg aacatggctg agctggccca
     2401 gcaggtggtg aagcaggagc tgcctagcga agagggccca ggggaggccc tgatgctggg
     2461 ggctgaggtc cctgaccctg agccactgcc agctctgccc ccgcaagccc cgctgcccct
     2521 gcccacccag ccaccgtccc cattccatca cctggacttc agtcacagcc tgagctttgg
     2581 gggcagggag gacgagggtc ccccgggcta ccccgaaccc ctggcaccgg ggcatggctc
     2641 cccattcccc agcctgtcca agaaggatct ggacctcatg ctcctggacg actcactgct
     2701 accgctggcc tctgatccac ttctgtccac catgtccccc gaggcctcca aggccagcag
     2761 ccgccggagc agcttcagca tggaggaggg cgatgtgctg gggcggccgc ccatggtgag
     2821 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2881 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     2941 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3001 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     3061 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3121 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3181 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     3241 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     3301 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     3361 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     3421 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     3481 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagatggccg
     3541 ctacgtaaat tccgcccccc cccccctctc ctcccccccc cctaacgtta ctggccgaag
     3601 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     3661 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     3721 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     3781 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     3841 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     3901 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     3961 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     4021 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg
     4081 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg
     4141 ccacaaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc
     4201 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     4261 acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     4321 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     4381 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     4441 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     4501 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     4561 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     4621 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     4681 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     4741 gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg
     4801 gctcgacgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     4861 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     4921 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
     4981 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     5041 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     5101 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     5161 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     5221 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     5281 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     5341 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     5401 ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa
     5461 ttaatttggt tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc
     5521 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5581 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5641 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5701 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5761 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5821 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5881 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5941 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6001 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6061 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6121 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6181 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6241 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6301 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6361 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6421 atcttcacct agatcctttt gcggccggcc gcaaatcaat ctaaagtata tatgagtaaa
     6481 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6541 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6601 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6661 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6721 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6781 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6841 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6901 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6961 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7021 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7081 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     7141 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7201 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7261 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7321 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7381 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7441 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     7501 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcact
     7561 cgaggagctt ggcccattgc atacgttgta tccatatcat aatatgtaca tttatattgg
     7621 ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc
     7681 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     7741 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     7801 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     7861 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     7921 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     7981 tcctacttgg cagtacatct acgtattagt catcgctatt accatgctag agaaccatca
     8041 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     8101 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     8161 acaacccctc actcggcgcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta
     8221 tccaataaac cctcttgcag ttgcatccga cttgtgg
//
