LOCUS       sequence-340904-        5549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses rtTA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5549)
  AUTHORS   Aihara S, Fujimoto S, Sakaguchi R, Imai T
  TITLE     BMPR-2 gates activity-dependent stabilization of primary dendrites 
            during mitral cell remodeling.
  JOURNAL   Cell Rep. 2021 Jun 22;35(12):109276. doi: 
            10.1016/j.celrep.2021.109276.
  PUBMED    34161760
REFERENCE   2  (bases 1 to 5549)
  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: "Cell Rep.";
            date: "2021-06-22"; volume: "35(12):109276. doi"; pages: " 
            10.1016/j.celrep.2021.109276"
COMMENT     Sequence Label: pCAG-rtTA
FEATURES             Location/Qualifiers
     source          1..5549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1739..2356,2357..2362,2363..2398,2399..2401,2402..2437,
                     2438..2440,2441..2476,2477..2485)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(2571..2590)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2636..2691
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2690..2709)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3052..3068)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3052..3068)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3065..3087)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3076..3092
                     /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(3100..3106,3107..3124,3125..3130))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3145..3166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3225..3420
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3271..3406
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3426..3560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3463..3482)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3517..3536
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3628..3645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3799..4387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3879..3898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4558..5349,5350..5418))
                     /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     5181..5200
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5419..5523)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccrggsggg gsggggsggg gsgrggggsg
      541 gggsggggsg rggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccaccat
     1741 gtctagactg gacaagagca aagtcataaa cggcgctctg gaattactca atggagtcgg
     1801 tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct
     1861 gtactggcac gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg agatgctgga
     1921 caggcatcat acccacttct gccccctgga aggcgagtca tggcaagact ttctgcggaa
     1981 caacgccaag tcattccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct
     2041 cggcacccgc ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg
     2101 tcagcaaggc ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac
     2161 actgggctgc gtattggagg aacaggagca tcaagtagca aaagaggaaa gagagacacc
     2221 taccaccgat tctatgcccc cacttctgag acaagcaatt gagctgttcg accggcaggg
     2281 agccgaacct gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct
     2341 aaagtgcgaa agcggcgggc cggccgacgc ccttgacgat tttgacttag acatgctccc
     2401 agccgatgcc cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt
     2461 tgaccttgac atgctccccg ggtaagcggc cgcactcctc aggtgcaggc tgcctatcag
     2521 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     2581 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     2641 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     2701 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     2761 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
     2821 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     2881 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     2941 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     3001 cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
     3061 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3121 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3181 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
     3241 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3301 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3361 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3421 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3481 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3541 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3601 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3661 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3721 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3781 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3841 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3901 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3961 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4021 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4081 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4141 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4201 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4261 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4321 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4381 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4441 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4501 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4561 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4621 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4681 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4741 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4801 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4861 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4921 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4981 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5041 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5101 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5161 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5221 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5281 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5341 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5401 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5461 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5521 gcgcacattt ccccgaaaag tgccacctg
//
