LOCUS       sequence-189314-        4973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox-inducible expression of Gsx2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4973)
  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 4973)
  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
FEATURES             Location/Qualifiers
     source          1..4973
                     /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"
     intron          2122..2187
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2317..2337
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2762..2883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2799..2818)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2853..2872
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2969..2986)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3140..3728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3220..3239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3899..4690,4691..4759))
                     /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     4522..4541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4760..4864)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4932..4950
                     /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 atggctagcc
     1981 tgcagggatc caatgtaact gtattcagcg atgacgaaat tcttagctat tgtaatactc
     2041 tagaggatct ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct
     2101 acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta
     2161 ctgattctaa ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag
     2221 tggtggaatg cctttaatga ggaaaacctg ttttgctcag aagaaatgcc atctagtgat
     2281 gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa
     2341 gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat
     2401 agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag
     2461 aaaattatgg aaaaatattc tgtaaccttt ataagtaggc ataacagtta taatcataac
     2521 atactgtttt ttcttactcc acacaggcat agagtgtctg ctattaataa ctatgctcaa
     2581 aaattgtgta cctttagctt tttaatttgt aaaggggtta ataaggaata tttgatgtat
     2641 agtgccttga ctagagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2701 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2761 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2821 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2881 ttatcatgtc tgcggctcta gagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2941 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3001 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3061 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3121 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3181 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3241 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3301 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3361 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3421 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3481 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3541 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3601 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3661 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3721 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3781 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3841 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3901 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3961 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4021 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4081 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4141 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4201 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4261 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4321 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4381 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4441 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4501 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4561 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4621 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4681 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4741 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4801 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4861 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     4921 taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaattc ctc
//
