LOCUS       sequence-266809-        4478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains the TetOn3G-transactivator sequence under control 
            of a Camk2a promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4478)
  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 4478)
  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: pCAMKIIa-TetOn3G
FEATURES             Location/Qualifiers
     source          1..4478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     595..614
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        595..613
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             667..1413
                     /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(1426..1443)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1432..1656
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1741..1881
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1741..1870
                     /label=AAV2 ITR
     rep_origin      1956..2411
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2043..2062)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2253..2274
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2428..2447)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2547..2569
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2607..2625)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2693..2797
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2798..2866,2867..3658)
                     /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(3016..3035)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3829..4417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4318..4337
                     /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 cttaggatgt
      241 ggactaagtt tgttcacatc cccttctcca accccctcag tacatcaccc tgggggaaca
      301 gggtccactt gctcctgggc ccacacagtc ctgcagtatt gtgtatataa ggccagggca
      361 aagaggagca ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt taccggggca
      421 acgggaacag ggcgtttcgg aggtggttgc catggggacc tggatgctga cgaaggctcg
      481 cgaggctgtg agcagccaca gtgccctgct cagaagcccc aagctcgtca gtcaagccgg
      541 ttctccgttt gcactcagga gcacgggcag gcgagtggcc cctagttctg ggagtaatac
      601 gactcactat agggagaccc aagctggcta gcgtttaaac ttaagcttgg taccgagctc
      661 ggatccatgt ctagactgga caagagcaaa gtcataaact ctgctctgga attactcaat
      721 ggagtcggta tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag
      781 cctaccctgt actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag
      841 atgctggaca ggcatcatac ccactcctgc cccctggaag gcgagtcatg gcaagacttt
      901 ctgcggaaca acgccaagtc ataccgctgt gctctcctct cacatcgcga cggggctaaa
      961 gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg
     1021 ttcctgtgtc agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc
     1081 cactttacac tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga
     1141 gagacaccta ccaccgattc tatgccccca cttctgaaac aagcaattga gctgttcgac
     1201 cggcagggag ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag
     1261 aaacagctaa agtgcgaaag cggcgggccg accgacgccc ttgacgattt tgacttagac
     1321 atgctcccag ccgatgccct tgacgacttt gaccttgata tgctgcctgc tgacgctctt
     1381 gacgattttg accttgacat gctccccggg taacccgctg atcagcctcg actgtgcctt
     1441 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1501 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     1561 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     1621 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagat
     1681 cctctcttaa ggtagcatcg agatttaaat tagggataac agggtaatgg cgcgggccgc
     1741 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     1801 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     1861 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     1921 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     1981 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     2041 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2101 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2161 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2221 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2281 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     2341 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     2401 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2461 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     2521 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     2581 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     2641 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     2701 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2761 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     2821 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     2881 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     2941 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3001 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3061 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3121 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3181 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3241 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3301 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3361 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3421 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3481 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3541 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3601 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3661 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3721 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3781 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3841 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3901 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3961 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4021 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4081 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4141 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4201 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4261 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4321 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4381 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4441 acggttcctg gccttttgct ggccttttgc tcacatgt
//
