LOCUS       sequence-189322-        6873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox-inducible expression of Gsx2 and Ebf1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6873)
  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 6873)
  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-GSX2-IRES2-EBF1
FEATURES             Location/Qualifiers
     source          1..6873
                     /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"
     CDS             807..824
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    join(1362..1937,1938..1940,1941..1949)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1530..1547)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1757..1776
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2002..2025
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     intron          4022..4087
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4217..4237
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4662..4783
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4699..4718)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4753..4772
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4869..4886)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5040..5628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5120..5139)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5799..6590,6591..6659))
                     /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     6422..6441
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6660..6764)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6832..6850
                     /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 catcgacatg tcgcgctcct
      421 tctacgtgga ctcactcatc atcaaggact cctcacggcc cgcaccctcg ctgccggaat
      481 cgcacccagg gccggatttc ttcatcccgc tgggcatgcc gtccccgttg gtgatgtcag
      541 tgtccgggcc tggctgtccg tcccgcaaaa gcggcgcttt ctgcgtgtgc ccgctttgcg
      601 tcacctcgca cctgcactcc tctcgcccgc ccgcaggagc tggtggcgga gctacgggga
      661 ccgcgggggc tgcagtggcg ggcggcgggg tggctggagg caccggcgct ctacctttgc
      721 tcaaaagcca gttctctccc gctcctgggg acgcgcagtt ttgcccacgg gtgagccacg
      781 cacaccatca ccaccaccca ccacagcatc atcatcacca tcaccagccc cagcagcccg
      841 gctcggctgc agcggcggcc gcggcggcgg cggcggcagc tgcggcagcg gcggctctgg
      901 gacacccgca gcaccacgca cctgtctgcg ccgccactac ctacaacatg tcggacccac
      961 ggagattcca ctgcctctcc atgggaggct ccgataccag ccaggtaccc aatggcaaaa
     1021 ggatgaggac agcgtttacc agcacgcagc tcctggagct ggagcgagaa ttctcttcca
     1081 atatgtacct gtcccgactc cggagaatcg agatcgcgac atacctaaac ctgtcagaga
     1141 agcaggtgaa aatctggttt cagaaccgtc gcgtgaagca caagaaggag gggaaaggcg
     1201 cttcgaggaa caatcacaca agctgcaagt gcgtgggtag ccaggcgcat tatgcgcgct
     1261 ccgaggacga ggactccttg tcccctgcct cggctaacga agacaaggag atttccccct
     1321 tgtaaaggca gaggctcctt ctggatctat cgatcggccg gcccctctcc ctcccccccc
     1381 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     1441 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     1501 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     1561 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     1621 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     1681 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     1741 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     1801 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct ttacatgtgt
     1861 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     1921 aaaaacacga tgataatatg gccacaaccg ggccggatat cacgcgtcat atggctagcg
     1981 tttaaactta agctaaccat ggactacaag gacgacgatg acaagggggt acctaggatc
     2041 cagtgtggtg gaattctgca gatatccagc acagtggcgg ccgctcgagc aatgtttggg
     2101 atccaggaaa gcatccaacg gagtggaagc agtatgaagg aagagccgct gggcagcggc
     2161 atgaacgcgg tgcggacgtg gatgcagggc gccggggtgc tggacgccaa cacagcggcg
     2221 cagagcgggg tgggtctggc ccgggctcac tttgagaagc agccgccttc taacctgcgg
     2281 aaatccaact tcttccactt cgtcctggcc ctctacgaca gacagggcca gcccgtggag
     2341 attgagagga ctgcctttgt ggggttcgtg gagaaggaaa aagaagccaa cagcgaaaag
     2401 accaataatg ggatccacta ccggctccag ctcctctaca gcaatgggat acggacagaa
     2461 caggatttct atgtgcgcct catcgactcc atgacaaaac aagccatagt gtatgaaggc
     2521 caagacaaga accctgaaat gtgccgagta ttgctcacac acgagatcat gtgcagccgc
     2581 tgttgtgaca agaaaagctg tggcaaccga aatgagactc cctcagatcc agtgataatt
     2641 gacaggttct tcctgaagtt tttccttaaa tgcaaccaaa attgcctaaa gaatgcagga
     2701 aacccacgtg acatgcggag attccaggtc gtggtgtcta ccacagtcaa cgtggatggc
     2761 catgtcctgg cagtctctga taacatgttt gtccacaata actccaagca cgggcggagg
     2821 gctcggaggc ttgacccctc ggaaggtacg ccctcttatc tggaacatgc tactccctgt
     2881 atcaaagcca tcagcccgag tgaaggatgg acgacgggag gcgcgactgt gatcatcata
     2941 ggggacaatt tctttgatgg gttacaggtc atattcggta ccatgctggt ctggagtgag
     3001 ttgataactc ctcatgccat ccgagttcag acacctcctc ggcacatccc tggtgtggtg
     3061 gaagtcacac tgtcgtacaa gtccaagcag ttctgcaaag ggacaccagg cagattcatc
     3121 tacacagcac tcaatgaacc caccatcgac tacggcttcc agaggttaca gaaggtcatt
     3181 cctcggcatc ctggtgaccc agagcgcttg ccaaaggaag tgatccttaa aagagctgca
     3241 gatctggttg aagccctgta tgggatgccc cacaacaacc aggagattat cctgaagaga
     3301 gctgccgaca ttgcagaggc tctgtacagt gtccctcgga accacaacca gctcccagcc
     3361 cttgctaaca cttcggtcca tgcagggatg atgggtgtga actccttcag tggacaactg
     3421 gctgtgaatg tctcggaggc atcacaagcc accaatcaag gtttcacccg caactcaagc
     3481 agcgtgtcac cacatggcta tgtgccgagc accaccccac agcaaaccaa ctataactca
     3541 gtcaccacaa gcatgaatgg ctacggctca gccgccatgt ccaatttggg cggctcccca
     3601 accttcctca atggctcagc tgccaactca ccctatgcca ttgtgccatc cagccccacc
     3661 atggcctcat ctacaagcct cccctccaac tgcagtagct cctctggcat cttctccttc
     3721 tcaccagcca acatggtctc agcagtgaaa cagaagagtg ctttcgcacc agttgtcaga
     3781 ccccagacgt cccctcctcc cacctgcacc agcaccaacg ggaacagcct gcaagcgata
     3841 tctggcatga ttgtccctcc catgtgactc gagtctagcc tgcagggatc caatgtaact
     3901 gtattcagcg atgacgaaat tcttagctat tgtaatactc tagaggatct ttgtgaagga
     3961 accttacttc tgtggtgtga cataattgga caaactacct acagagattt aaagctctaa
     4021 ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt
     4081 attttagatt ccaacctatg gaactgatga atgggagcag tggtggaatg cctttaatga
     4141 ggaaaacctg ttttgctcag aagaaatgcc atctagtgat gatgaggcta ctgctgactc
     4201 tcaacattct actcctccaa aaaagaagag aaaggtagaa gaccccaagg actttccttc
     4261 agaattgcta agttttttga gtcatgctgt gtttagtaat agaactcttg cttgctttgc
     4321 tatttacacc acaaaggaaa aagctgcact gctatacaag aaaattatgg aaaaatattc
     4381 tgtaaccttt ataagtaggc ataacagtta taatcataac atactgtttt ttcttactcc
     4441 acacaggcat agagtgtctg ctattaataa ctatgctcaa aaattgtgta cctttagctt
     4501 tttaatttgt aaaggggtta ataaggaata tttgatgtat agtgccttga ctagagatca
     4561 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     4621 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     4681 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4741 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgcggctcta
     4801 gagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4861 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4921 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4981 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5041 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5101 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5161 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5221 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5281 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5341 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5401 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5461 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5521 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5581 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5641 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5701 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5761 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5821 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5881 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5941 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6001 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6061 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6121 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6181 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6241 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6301 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6361 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6421 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6481 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6541 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6601 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6661 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6721 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6781 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     6841 atcacgaggc cctttcgtct tcaagaattc ctc
//
