LOCUS       sequence-256995-        5810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains the TetOn3G-transactivator sequence under control 
            of a GFAP promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5810)
  AUTHORS   Heumuller M, Glock C, Rangaraju V, Biever A, Schuman EM
  TITLE     A genetically encodable cell-type-specific protein synthesis 
            inhibitor.
  JOURNAL   Nat Methods. 2019 Jul 15. pii: 10.1038/s41592-019-0468-x. doi: 
            10.1038/s41592-019-0468-x.
  PUBMED    31308551
REFERENCE   2  (bases 1 to 5810)
  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"; journalName: "Nat 
            Methods. 2019 Jul 15. pii: 10.1038/s41592-019-0468-x. doi: 
            10.1038/s41592-019-0468-x."
COMMENT     Sequence Label: pGFAP-TetOn3G
FEATURES             Location/Qualifiers
     source          1..5810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     CDS             1999..2745
                     /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"
     primer_bind     complement(2758..2775)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2764..2988
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3073..3213
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3073..3202
                     /label=AAV2 ITR
     rep_origin      3288..3743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3375..3394)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3585..3606
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3760..3779)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3879..3901
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3939..3957)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4025..4129
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4130..4198,4199..4990)
                     /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     complement(4348..4367)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5161..5749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5650..5669
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccaat tcagtcgata actataacgg tcctaaggta
      181 gcgatttaaa tacgcgctct cttaaggtag ccccgggacg cgtcaattgc atgaagaatc
      241 tgcttagggt ctgtaagctg aagacctggc agtgctgagc tggtcagccc ccaggacctc
      301 cttttgtgcc cacgagtgaa tcaccttggc atagacataa tggtcagggg tgggcacgca
      361 gcctgcttcc cgctgtgctc caggcctcct tcgatgcttt ccgagaagtc tattgagctg
      421 ggagcttgta ctgcacccgg ggctgacatc ctggcatcct gggataaaag cagcccacgg
      481 ggctgccctt gccatatgcc tcactggcgg cagagaacaa ggctctattc agcgagtacc
      541 ctggagtaga caccagaagc ccaagcatgg gcagaggaag gcaggggttg gggggagcag
      601 agctgtctgt gttccagaag cccaaggaca cagatggcta aggcgcctgg gagagggacc
      661 tgagtggaag agatagatgg gcctgaagtc tcaagcagca acagcctcct ccccgccatt
      721 ggtgagggtg gggtttggtt tcccggacct acatatccct cagaggcctg gtgtgtagga
      781 atttaaaggg ggtaaatctc ctgagagaat gaggggtacc caggaagacg gggtgttaca
      841 gaaagaaaga ctccagcatg cacagccaac tcattcaaaa ctactctgtc aggggctgcc
      901 aggggccagg ctcggggtgg ggggtggggg gcaacgagaa gctggatcag ggagaaatgg
      961 cccactaggc tggataagag gccacagagg ggctcaggaa tgaagcctgc tgtcttaccc
     1021 tattaggatc tgcgtgcata ccttctgccg tgcactctaa acacacagcc agaggctcaa
     1081 gttgaccctg gagtcacaga gagggctcca accttagccc tccactcctg aactccagga
     1141 atgagaagat agagttggag agattcaggg gagaggactc tgttgagaat gggggtcaca
     1201 ggaaactgta atataggttg atcccggagg aagggaatag gttcttcaag ttcctagcat
     1261 ctcacaggcc cccagagaag gacagagttg gggtggtcct ggcttacagg ctctaagaac
     1321 tggaagctga ttaccccacc gagctgtgca ctctctgtct ctgtctctgt gtgtgcgctc
     1381 gtgcacactt atcacacaaa tgttcatgtg tgtgcacata catgtgttga gaccagaggt
     1441 caacctcagg cactgttgcc ttggttttct gagagagcat ttctctctgg atctggaact
     1501 cgccaattag tgagagccag gaagtctgct gattttcact gcccagcact ggagtttaca
     1561 agtatgcact gtcaacccag gccttttgta ttcattctgc agctagaact tgggtgggtc
     1621 ttcatgcttg acaggcaagc aatttatgga ctaagctgtt ccctcggccc tctcttgacc
     1681 catttaccag aaagggggtt ccttgatcaa tggcgaagcc aggctggtgt tcccaagaaa
     1741 gccttgactc tgggtacagt gacctcagtg gggtgagagg agttctcccc ctagctgggc
     1801 tggggcccag ctccaccccc tcaggctatt caatgggggt gcttccagga agtcaggggc
     1861 agatttagtc caacccgttc ctccataaag gccctgacat cccaggagcc agcagaggca
     1921 gggcaccaat acgactcact atagggagac ccaagctggc tagcgtttaa acttaagctt
     1981 ggtaccgagc tcggatccat gtctagactg gacaagagca aagtcataaa ctctgctctg
     2041 gaattactca atggagtcgg tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg
     2101 ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct gctcgatgcc
     2161 ctgccaatcg agatgctgga caggcatcat acccactcct gccccctgga aggcgagtca
     2221 tggcaagact ttctgcggaa caacgccaag tcataccgct gtgctctcct ctcacatcgc
     2281 gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga aaccctggaa
     2341 aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact gtacgctctg
     2401 tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca tcaagtagca
     2461 aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgaa acaagcaatt
     2521 gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga actaatcata
     2581 tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cgaccgacgc ccttgacgat
     2641 tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga tatgctgcct
     2701 gctgacgctc ttgacgattt tgaccttgac atgctccccg ggtaacccgc tgatcagcct
     2761 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2821 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2881 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2941 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     3001 aaagaaccag atcctctctt aaggtagcat cgagatttaa attagggata acagggtaat
     3061 ggcgcgggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3121 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3181 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     3241 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3301 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3361 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3421 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3481 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3541 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3601 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc
     3661 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3721 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3781 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3841 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3901 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3961 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4021 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4081 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4141 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4201 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4261 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4321 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4381 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4441 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4501 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4561 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4621 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4681 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4741 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4801 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4861 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4921 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4981 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5041 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5101 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5161 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5221 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5281 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     5341 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5401 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5461 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5521 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5581 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5641 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5701 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5761 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
