LOCUS       sequence-198128-        6921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6921)
  AUTHORS   Vega-Rubin-de-Celis S, Pena-Llopis S, Konda M, Brugarolas J
  TITLE     Multistep regulation of TFEB by MTORC1.
  JOURNAL   Autophagy. 2017 Mar 4;13(3):464-472. doi: 
            10.1080/15548627.2016.1271514. Epub 2017 Jan 5.
  PUBMED    28055300
REFERENCE   2  (bases 1 to 6921)
  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: 
            "10.1080/15548627.2016.1271514"; journalName: "Autophagy"; date: 
            "2017-03-4- 4"; volume: "13"; issue: "3"; pages: "464-472"
COMMENT     Sequence Label: pcDNA3.1-TFEB-WT-MYC (#647)
FEATURES             Location/Qualifiers
     source          1..6921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      726..735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2172..2201
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2217..2234
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2278..2295)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2284..2508
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2554..2982
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2641..2660)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2851..2872
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2991..3011)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2996..3325
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3176..3311
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3238..3257
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3392..4186
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3446..3465)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4056..4075
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4360..4481
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4397..4416)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4451..4470
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4530..4546)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4530..4546)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4543..4565)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4554..4570
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4578..4584,4585..4602,4603..4608))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4623..4644
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4761..4778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4932..5520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5012..5031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5691..6482,6483..6551))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6314..6333
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6552..6656)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6731..6750)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacgggccc tctagactcg agcggccgcc actgtgctgg atatctgcag
      721 aattcgccac catggcgtca cgcatagggt tgcgcatgca gctcatgcgg gagcaggcgc
      781 agcaggagga gcaacgggag cgcatgcagc aacaggctgt catgcattac atgcagcagc
      841 agcagcagca gcaacagcag cagctcggag ggccgcccac cccggccatc aatacccccg
      901 tccacttcca gtcgccacca cctgtgcctg gggaggtgtt gaaggtgcag tcctacctgg
      961 agaatcccac atcctaccat ctgcagcagt cgcagcatca gaaggtgcgg gagtacctgt
     1021 ccgagaccta tggaaacaag tttgctgccc acatcagccc agcccagggc tctccgaaac
     1081 ccccaccagc cgcctcccca ggggtgcgag ctggacacgt gctgtcctcc tccgctggca
     1141 acagtgctcc caatagcccc atggccatgc tgcacattgg ctccaaccct gagagggagt
     1201 tggatgatgt cattgacaac attatgcgtc tggacgatgt ccttggctac atcaatcctg
     1261 aaatgcagat gcccaacacg ctacccctgt ccagcagcca cctgaatgtg tacagcagcg
     1321 acccccaggt cacagcctcc ctggtgggcg tcaccagcag ctcctgccct gcggacctga
     1381 cccagaagcg agagctcaca gatgctgaga gcagggccct ggccaaggag cggcagaaga
     1441 aagacaatca caacttaatt gaaaggagac gaaggttcaa catcaatgac cgcatcaagg
     1501 agttgggaat gctgatcccc aaggccaatg acctggacgt gcgctggaac aagggcacca
     1561 tcctcaaggc ctctgtggat tacatccgga ggatgcagaa ggacctgcaa aagtccaggg
     1621 agctggagaa ccactctcgc cgcctggaga tgaccaacaa gcagctctgg ctccgtatcc
     1681 aggagctgga gatgcaggct cgagtgcacg gcctccctac cacctccccg tccggcatga
     1741 acatggctga gctggcccag caggtggtga agcaggagct gcctagcgaa gagggcccag
     1801 gggaggccct gatgctgggg gctgaggtcc ctgaccctga gccactgcca gctctgcccc
     1861 cgcaagcccc gctgcccctg cccacccagc caccgtcccc attccatcac ctggacttca
     1921 gccacagcct gagctttggg ggcagggagg acgagggtcc cccgggctac cccgaacccc
     1981 tggcgccggg gcatggctcc ccattcccca gcctgtccaa gaaggatctg gacctcatgc
     2041 tcctggacga ctcactgcta ccgctggcct ctgatccact tctgtccacc atgtcccccg
     2101 aggcctccaa ggccagcagc cgccggagca gcttcagcat ggaggagggc gatgtgctgt
     2161 caagctttct agaacaaaaa ctcatctcag aagaggatct gaatagcgcc gtcgaccatc
     2221 atcatcatca tcattgagtt taaacggtct ccagcttaag tttaaaccgc tgatcagcct
     2281 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2341 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2401 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2461 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     2521 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2581 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2641 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2701 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2761 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2821 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2881 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2941 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3001 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3061 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3121 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3181 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3241 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3301 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3361 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3421 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3481 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3541 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3601 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3661 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3721 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3781 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3841 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3901 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3961 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4021 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4081 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4141 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4201 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4261 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4321 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4381 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4441 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4501 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4561 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4621 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4681 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4741 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4801 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4861 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4921 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4981 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5041 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5101 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5161 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5221 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5281 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5341 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5401 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5461 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5521 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5581 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5641 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5701 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5761 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5821 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5881 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5941 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6001 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6061 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6121 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6181 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6241 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6301 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6361 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6421 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6481 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6541 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6601 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6661 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     6721 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     6781 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     6841 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     6901 tacgggccag atatacgcgt t
//
