LOCUS       sequence-189317-        5920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox-inducible expression of Ebf1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5920)
  AUTHORS   Faedo A, Laporta A, Segnali A, Galimberti M, Besusso D, Cesana E, 
            Belloli S, Moresco RM, Tropiano M, Fuca E, Wild S, Bosio A, Vercelli
            AE, Biella G, Cattaneo E
  TITLE     Differentiation of human telencephalic progenitor cells into MSNs by
            inducible expression of Gsx2 and Ebf1.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Feb 14;114(7):E1234-E1242. doi: 
            10.1073/pnas.1611473114. Epub 2017 Jan 30.
  PUBMED    28137879
REFERENCE   2  (bases 1 to 5920)
  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: 
            "10.1073/pnas.1611473114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2017-02-14- 14"; volume: "114"; issue: "7"; pages: 
            "E1234-E1242"
COMMENT     Sequence Label: pTRE3G-EBF1
FEATURES             Location/Qualifiers
     source          1..5920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..379
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    9..27
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    45..63
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    81..99
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    117..135
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    153..171
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    189..207
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    225..243
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     288..312
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    join(415..990,991..993,994..1002)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(583..600)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     810..829
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1055..1078
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     intron          3069..3134
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3264..3284
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3709..3830
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3746..3765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3800..3819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3916..3933)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4087..4675)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4167..4186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4846..5637,5638..5706))
                     /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     5469..5488
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5707..5811)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5879..5897
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta tcagtgatag
       61 agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga gtttactccc
      121 tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag aacgtatcta
      181 cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta tcagtgatag
      241 agaacgtata agctttaggc gtgtacggtg ggcgcctata aaagcagagc tcgtttagtg
      301 aaccgtcaga tcgcctggag caattccaca acacttttgt cttataccaa ctttccgtac
      361 cacttcctac cctcgtaaag tcgacaccgg ggcccagatc tatcgatcgg ccggcccctc
      421 tccctccccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
      481 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
      541 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
      601 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
      661 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
      721 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
      781 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
      841 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtacacat
      901 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
      961 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccgggccgga tatcacgcgt
     1021 catatggcta gcgtttaaac ttaagctaac catggactac aaggacgacg atgacaaggg
     1081 ggtacctagg atccagtgtg gtggaattct gcagatatcc agcacagtgg cggccgctcg
     1141 agcaatgttt gggatccagg aaagcatcca acggagtgga agcagtatga aggaagagcc
     1201 gctgggcagc ggcatgaacg cggtgcggac gtggatgcag ggcgccgggg tgctggacgc
     1261 caacacagcg gcgcagagcg gggtgggtct ggcccgggct cactttgaga agcagccgcc
     1321 ttctaacctg cggaaatcca acttcttcca cttcgtcctg gccctctacg acagacaggg
     1381 ccagcccgtg gagattgaga ggactgcctt tgtggggttc gtggagaagg aaaaagaagc
     1441 caacagcgaa aagaccaata atgggatcca ctaccggctc cagctcctct acagcaatgg
     1501 gatacggaca gaacaggatt tctatgtgcg cctcatcgac tccatgacaa aacaagccat
     1561 agtgtatgaa ggccaagaca agaaccctga aatgtgccga gtattgctca cacacgagat
     1621 catgtgcagc cgctgttgtg acaagaaaag ctgtggcaac cgaaatgaga ctccctcaga
     1681 tccagtgata attgacaggt tcttcctgaa gtttttcctt aaatgcaacc aaaattgcct
     1741 aaagaatgca ggaaacccac gtgacatgcg gagattccag gtcgtggtgt ctaccacagt
     1801 caacgtggat ggccatgtcc tggcagtctc tgataacatg tttgtccaca ataactccaa
     1861 gcacgggcgg agggctcgga ggcttgaccc ctcggaaggt acgccctctt atctggaaca
     1921 tgctactccc tgtatcaaag ccatcagccc gagtgaagga tggacgacgg gaggcgcgac
     1981 tgtgatcatc ataggggaca atttctttga tgggttacag gtcatattcg gtaccatgct
     2041 ggtctggagt gagttgataa ctcctcatgc catccgagtt cagacacctc ctcggcacat
     2101 ccctggtgtg gtggaagtca cactgtcgta caagtccaag cagttctgca aagggacacc
     2161 aggcagattc atctacacag cactcaatga acccaccatc gactacggct tccagaggtt
     2221 acagaaggtc attcctcggc atcctggtga cccagagcgc ttgccaaagg aagtgatcct
     2281 taaaagagct gcagatctgg ttgaagccct gtatgggatg ccccacaaca accaggagat
     2341 tatcctgaag agagctgccg acattgcaga ggctctgtac agtgtccctc ggaaccacaa
     2401 ccagctccca gcccttgcta acacttcggt ccatgcaggg atgatgggtg tgaactcctt
     2461 cagtggacaa ctggctgtga atgtctcgga ggcatcacaa gccaccaatc aaggtttcac
     2521 ccgcaactca agcagcgtgt caccacatgg ctatgtgccg agcaccaccc cacagcaaac
     2581 caactataac tcagtcacca caagcatgaa tggctacggc tcagccgcca tgtccaattt
     2641 gggcggctcc ccaaccttcc tcaatggctc agctgccaac tcaccctatg ccattgtgcc
     2701 atccagcccc accatggcct catctacaag cctcccctcc aactgcagta gctcctctgg
     2761 catcttctcc ttctcaccag ccaacatggt ctcagcagtg aaacagaaga gtgctttcgc
     2821 accagttgtc agaccccaga cgtcccctcc tcccacctgc accagcacca acgggaacag
     2881 cctgcaagcg atatctggca tgattgtccc tcccatgtga ctcgagtcta gaggatccaa
     2941 tgtaactgta ttcagcgatg acgaaattct tagctattgt aatactctag aggatctttg
     3001 tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa
     3061 gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg
     3121 tttgtgtatt ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct
     3181 ttaatgagga aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg
     3241 ctgactctca acattctact cctccaaaaa agaagagaaa ggtagaagac cccaaggact
     3301 ttccttcaga attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt
     3361 gctttgctat ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa
     3421 aatattctgt aacctttata agtaggcata acagttataa tcataacata ctgttttttc
     3481 ttactccaca caggcataga gtgtctgcta ttaataacta tgctcaaaaa ttgtgtacct
     3541 ttagcttttt aatttgtaaa ggggttaata aggaatattt gatgtatagt gccttgacta
     3601 gagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3661 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3721 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3781 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgc
     3841 ggctctagag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3901 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3961 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4021 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4081 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4141 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4201 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4261 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4321 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4381 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4441 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4501 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4561 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4621 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4681 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4741 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4801 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4861 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4921 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4981 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5041 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5101 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5161 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5221 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5281 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5341 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5401 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5461 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5521 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5581 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5641 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5701 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5761 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5821 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     5881 taggcgtatc acgaggccct ttcgtcttca agaattcctc
//
