LOCUS       sequence-208142-        4658 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses tTA2 under the CAG promoter in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4658)
  AUTHORS   Sakaguchi R, Leiwe MN, Imai T
  TITLE     Bright multicolor labeling of neuronal circuits with fluorescent 
            proteins and chemical tags.
  JOURNAL   eLife. 2018 Nov 20;7. pii: 40350. doi: 10.7554/eLife.40350.
  PUBMED    30454553
REFERENCE   2  (bases 1 to 4658)
  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: "eLife."; 
            date: "2018-11-20"; volume: "7. pii: 40350. doi"; pages: " 
            10.7554/eLife.40350"
COMMENT     Sequence Label: pCAG-tTA
FEATURES             Location/Qualifiers
     source          1..4658
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     38..60
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     122..141
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(175..792,793..798,799..834,835..837,838..873,874..876,
                     877..912,913..921)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=tTA-Advanced
                     /note="In the Tet-Off(R) system, tTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(1007..1026)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1072..1127
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1126..1145)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1488..1504)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1488..1504)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1501..1523)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1512..1528
                     /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(1536..1542,1543..1560,1561..1566))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1581..1602
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1661..1856
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1707..1842
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1862..1996
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1899..1918)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1953..1972
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2064..2081)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2235..2823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2315..2334)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2994..3785,3786..3854))
                     /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     3617..3636
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3855..3959)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3990..4369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4371..4646
                     /label=chicken beta-actin promoter
ORIGIN
        1 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
       61 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
      121 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgc caccatgtct
      181 agactggaca agagcaaagt cataaactct gctctggaat tactcaatga agtcggtatc
      241 gaaggcctga cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac
      301 tggcacgtga agaacaagcg ggccctgctc gatgccctgg caatcgagat gctggacagg
      361 catcataccc acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac
      421 gccaagtcat tccgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc
      481 acccgcccaa cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag
      541 caaggcttct ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg
      601 ggctgcgtat tggaggatca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc
      661 accgattcta tgcccccact tctgagacaa gcaattgagc tgttcgacca tcagggagcc
      721 gaacctgcct tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag
      781 tgcgaaagcg gcgggccggc cgacgccctt gacgattttg acttagacat gctcccagcc
      841 gatgcccttg acgactttga ccttgatatg ctgcctgctg acgctcttga cgattttgac
      901 cttgacatgc tccccgggta agcggccgca ctcctcaggt gcaggctgcc tatcagaagg
      961 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     1021 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     1081 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     1141 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     1201 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     1261 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     1321 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     1381 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     1441 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     1501 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     1561 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     1621 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
     1681 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     1741 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     1801 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     1861 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1921 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1981 ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2041 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2101 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2161 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2221 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2281 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2341 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2401 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2461 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2521 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2581 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2641 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2701 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2761 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2821 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2881 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2941 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3001 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3061 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3121 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3181 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3241 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3301 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3361 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3421 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3481 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3541 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3601 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3661 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3721 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3781 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3841 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3901 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3961 acatttcccc gaaaagtgcc acctgggtcg acattgatta ttgactagtt attaatagta
     4021 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     4081 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     4141 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     4201 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     4261 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     4321 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
     4381 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
     4441 atttattttt taattatttt gtgcagcgrt gggggcgggg gggggggggg cgcgcgccrg
     4501 gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca
     4561 atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct
     4621 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcg
//
