LOCUS       sequence-189783-        4702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Clontech rtTA driven by human beta-actin 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4702)
  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 4702)
  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: pCAG-TetON-3G
FEATURES             Location/Qualifiers
     source          1..4702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     830..852
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     914..933
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             964..1710
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    1720..1854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2205..2221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2205..2221)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2218..2240)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2229..2245
                     /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(2253..2259,2260..2277,2278..2283))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2298..2319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2378..2573
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2424..2559
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2579..2713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2781..2798)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2952..3540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3032..3051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3711..4502,4503..4571))
                     /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     4334..4353
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4572..4676)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg
      541 agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc
      601 gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc
      661 cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg
      721 ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg
      781 ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc
      841 tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc
      901 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc
      961 accatgtcta gactggacaa gagcaaagtc ataaactctg ctctggaatt actcaatgga
     1021 gtcggtatcg aaggcctgac gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct
     1081 accctgtact ggcacgtgaa gaacaagcgg gccctgctcg atgccctgcc aatcgagatg
     1141 ctggacaggc atcataccca ctcctgcccc ctggaaggcg agtcatggca agactttctg
     1201 cggaacaacg ccaagtcata ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg
     1261 catctcggca cccgcccaac agagaaacag tacgaaaccc tggaaaatca gctcgcgttc
     1321 ctgtgtcagc aaggcttctc cctggagaac gcactgtacg ctctgtccgc cgtgggccac
     1381 tttacactgg gctgcgtatt ggaggaacag gagcatcaag tagcaaaaga ggaaagagag
     1441 acacctacca ccgattctat gcccccactt ctgaaacaag caattgagct gttcgaccgg
     1501 cagggagccg aacctgcctt ccttttcggc ctggaactaa tcatatgtgg cctggagaaa
     1561 cagctaaagt gcgaaagcgg cgggccgacc gacgcccttg acgattttga cttagacatg
     1621 ctcccagccg atgcccttga cgactttgac cttgatatgc tgcctgctga cgctcttgac
     1681 gattttgacc ttgacatgct ccccgggtaa ctaagtaagg atccagacat gataagatac
     1741 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     1801 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     1861 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     1921 aagtaaaacc tctacaaatg tggtatggct gattatgatc ctgcaagcct cgtcgtcctg
     1981 gccggaccac gctatctgtg caaggtcccc ggccccggac gcgcgctcca tgagcagagc
     2041 gcccgccgcc gaggcgaaga ctcgggcggc gccctgcccg tcccaccagg tcaacaggcg
     2101 gtaaccggcc tcttcatcgg gaatgcgcgc gaccttcagc atcgccggca tgtccccctg
     2161 gcggacggga agtatccagc tcgaccaagc ttggcgtaat catggtcata gctgtttcct
     2221 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2281 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2341 gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca
     2401 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2461 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     2521 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa
     2581 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2641 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2701 tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2761 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2821 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2881 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2941 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3001 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3061 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3121 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3181 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3241 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3301 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3361 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3421 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3481 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3541 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3601 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3661 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3721 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3781 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3841 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3901 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3961 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4021 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4081 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4141 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4201 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4261 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4321 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4381 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4441 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4501 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4561 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4621 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4681 tttccccgaa aagtgccacc tg
//
