LOCUS       sequence-266812-        5798 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 5798)
  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 5798)
  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..5798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1987..2733
                     /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(2746..2763)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2752..2976
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3061..3201
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3061..3190
                     /label=AAV2 ITR
     rep_origin      3276..3731
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3363..3382)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3573..3594
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3748..3767)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3867..3889
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3927..3945)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4013..4117
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4118..4186,4187..4978)
                     /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(4336..4355)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5149..5737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5638..5657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggcaattc agtcgataac tataacggtc ctaaggtagc gatttaaata
      181 cgcgctctct taaggtagcc ccgggacgcg tcaattgcat gaagaatctg cttagggtct
      241 gtaagctgaa gacctggcag tgctgagctg gtcagccccc aggacctcct tttgtgccca
      301 cgagtgaatc accttggcat agacataatg gtcaggggtg ggcacgcagc ctgcttcccg
      361 ctgtgctcca ggcctccttc gatgctttcc gagaagtcta ttgagctggg agcttgtact
      421 gcacccgggg ctgacatcct ggcatcctgg gataaaagca gcccacgggg ctgcccttgc
      481 catatgcctc actggcggca gagaacaagg ctctattcag cgagtaccct ggagtagaca
      541 ccagaagccc aagcatgggc agaggaaggc aggggttggg gggagcagag ctgtctgtgt
      601 tccagaagcc caaggacaca gatggctaag gcgcctggga gagggacctg agtggaagag
      661 atagatgggc ctgaagtctc aagcagcaac agcctcctcc ccgccattgg tgagggtggg
      721 gtttggtttc ccggacctac atatccctca gaggcctggt gtgtaggaat ttaaaggggg
      781 taaatctcct gagagaatga ggggtaccca ggaagacggg gtgttacaga aagaaagact
      841 ccagcatgca cagccaactc attcaaaact actctgtcag gggctgccag gggccaggct
      901 cggggtgggg ggtggggggc aacgagaagc tggatcaggg agaaatggcc cactaggctg
      961 gataagaggc cacagagggg ctcaggaatg aagcctgctg tcttacccta ttaggatctg
     1021 cgtgcatacc ttctgccgtg cactctaaac acacagccag aggctcaagt tgaccctgga
     1081 gtcacagaga gggctccaac cttagccctc cactcctgaa ctccaggaat gagaagatag
     1141 agttggagag attcagggga gaggactctg ttgagaatgg gggtcacagg aaactgtaat
     1201 ataggttgat cccggaggaa gggaataggt tcttcaagtt cctagcatct cacaggcccc
     1261 cagagaagga cagagttggg gtggtcctgg cttacaggct ctaagaactg gaagctgatt
     1321 accccaccga gctgtgcact ctctgtctct gtctctgtgt gtgcgctcgt gcacacttat
     1381 cacacaaatg ttcatgtgtg tgcacataca tgtgttgaga ccagaggtca acctcaggca
     1441 ctgttgcctt ggttttctga gagagcattt ctctctggat ctggaactcg ccaattagtg
     1501 agagccagga agtctgctga ttttcactgc ccagcactgg agtttacaag tatgcactgt
     1561 caacccaggc cttttgtatt cattctgcag ctagaacttg ggtgggtctt catgcttgac
     1621 aggcaagcaa tttatggact aagctgttcc ctcggccctc tcttgaccca tttaccagaa
     1681 agggggttcc ttgatcaatg gcgaagccag gctggtgttc ccaagaaagc cttgactctg
     1741 ggtacagtga cctcagtggg gtgagaggag ttctacccct agctgggctg gggcccagct
     1801 ccaccccctc aggctattca atgggggtgc ttccaggaag tcaggggcag atttagtcca
     1861 acccgttcct ccataaaggc cctgacatcc caggagccag cagaggcagg gcaccaatac
     1921 gactcactat agggagaccc aagctggcta gcgtttaaac ttaagcttgg taccgagctc
     1981 ggatccatgt ctagactgga caagagcaaa gtcataaact ctgctctgga attactcaat
     2041 ggagtcggta tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag
     2101 cctaccctgt actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag
     2161 atgctggaca ggcatcatac ccactcctgc cccctggaag gcgagtcatg gcaagacttt
     2221 ctgcggaaca acgccaagtc ataccgctgt gctctcctct cacatcgcga cggggctaaa
     2281 gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg
     2341 ttcctgtgtc agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc
     2401 cactttacac tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga
     2461 gagacaccta ccaccgattc tatgccccca cttctgaaac aagcaattga gctgttcgac
     2521 cggcagggag ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag
     2581 aaacagctaa agtgcgaaag cggcgggccg accgacgccc ttgacgattt tgacttagac
     2641 atgctcccag ccgatgccct tgacgacttt gaccttgata tgctgcctgc tgacgctctt
     2701 gacgattttg accttgacat gctccccggg taacccgctg atcagcctcg actgtgcctt
     2761 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2821 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2881 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2941 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagat
     3001 cctctcttaa ggtagcatcg agatttaaat tagggataac agggtaatgg cgcgggccgc
     3061 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3121 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3181 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     3241 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3301 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     3361 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3421 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3481 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3541 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3601 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     3661 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3721 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3781 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3841 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3901 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3961 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4021 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4081 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4141 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4201 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4261 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4321 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4381 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4441 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4501 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4561 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4621 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4681 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4741 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4801 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4861 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4921 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4981 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5041 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5101 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5161 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5221 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5281 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5341 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5401 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5461 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5521 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5581 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5641 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5701 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5761 acggttcctg gccttttgct ggccttttgc tcacatgt
//
