LOCUS       sequence-155895-        6685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TetActivator protein under the control of the c-fos 
            promoter for use in TetTaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6685)
  AUTHORS   Zhang Z, Ferretti V, Guntan I, Moro A, Steinberg EA, Ye Z, Zecharia 
            AY, Yu X, Vyssotski AL, Brickley SG, Yustos R, Pillidge ZE, Harding 
            EC, Wisden W, Franks NP
  TITLE     Neuronal ensembles sufficient for recovery sleep and the sedative 
            actions of alpha2 adrenergic agonists.
  JOURNAL   Nat Neurosci. 2015 Apr;18(4):553-61. doi: 10.1038/nn.3957. Epub 2015
            Feb 23.
  PUBMED    25706476
REFERENCE   2  (bases 1 to 6685)
  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"; doi: "10.1038/nn.3957"; 
            journalName: "Nat Neurosci"; date: "2015-04"; volume: "18"; issue: 
            "4"; pages: "553-61"
COMMENT     Sequence Label: pAAV-cFos-tTA-pA
FEATURES             Location/Qualifiers
     source          1..6685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..52)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     152..171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     385..404
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        694..712
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     repeat_region   733..862
                     /label=AAV2 ITR
     CDS             903..956
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(1754..1765)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     intron          1823..2052
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2035..2054
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2722..2742
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2743..2766
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2900..2926
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2996..3014)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     intron          3098..3163
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     misc_feature    3216..3804
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3269..3289)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3687..3698)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3831..3848)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3837..4061
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4285..4420
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4606..4626
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4867..5455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4947..4966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5626..6417,6418..6486))
                     /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     6249..6268
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6487..6591)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6659..6677
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
       61 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
      121 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
      181 caccattcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
      241 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
      301 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
      361 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
      421 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatctggc tagcgatgac
      481 cctgctgatt ggttcgctga ccatttccgg gtgcgggacg gcgttaccag aaactcagaa
      541 ggttcgtcca accaaaccga ctctgacgac agtttacgat agagatgata gggtctgctt
      601 cagtaagcca gatgctacac aattaggctt gtacatattg tcgttagaac gcggctacaa
      661 ttaatacata accttatgta tcatacacat acgatttagg tgacactata gaatacacgg
      721 aattaattct agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
      781 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      841 aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta cttatctacc
      901 tcgagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc ggccctgcac
      961 cgggatccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1021 tcgagctgga cggttaatta agccagacgg ccgcagcttt cctttaggaa cagaggcttc
     1081 gagcctttaa ggctgcgtac ttgcttctcc taataccaga gactcaaaaa aaaaaaaaaa
     1141 gttccagatt gctggacaat gacccgggtc tcatcccttg accctgggaa ccgggtccac
     1201 attgaatcag gtgcgaatgt tcgctcgcct tctctgcctt tcccgcctcc cctcccccgg
     1261 ccgcggcccc ggttcccccc ctgcgctgca ccctcagagt tggctgcagc cggcgagctg
     1321 ttcccgtcaa tccctccctc ctttacacag gatgtccata ttaggacatc tgcgtcagca
     1381 ggtttccacg gccggtccct gttgttctgg gggggggacc atctccgaaa tcctacacgc
     1441 ggaaggtcta ggagaccccc taagatccca aatgtgaaca ctcataggtg aaagatgtat
     1501 gccaagacgg gggttgaaag cctggggcgt agagttgacg acagagcgcc cgcagagggc
     1561 cttggggcgc gcttcccccc ccttccagtt ccgcccagtg acgtaggaag tccatccatt
     1621 cacagcgctt ctataaaggc gccagctgag gcgcctacta ctccaaccgc gactgcagcg
     1681 agcaactgag aagactggat agagccggcg gttccgcgaa cgagcagtga ccgcgctccc
     1741 acccagctct gctctgcagc tcccaccagt gtctggccgc atcgattcta gaattcgctg
     1801 tctgcgaggg ccagctgttg gggtgagtac tccctctcaa aagcgggcat gacttctgcg
     1861 ctaagattgt cagtttccaa aaacgaggag gatttgatat tcacctggcc cgcggtgatg
     1921 cctttgaggg tggccgcgtc catctggtca gaaaagacaa tctttttgtt gtcaagcttg
     1981 aggtgtggca ggcttgagat ctggccatac acttgagtga caatgacatc cactttgcct
     2041 ttctctccac aggtgtccac tcccaggtcc aactgcagcc caagcggatc ggccgccacc
     2101 atgtcccgcc tggacaagtc caaggtgatc aactccgccc tggagctgct gaacgaggtg
     2161 ggcatcgagg gcctgaccac ccgcaagctg gcccagaagc tgggcgtgga gcagcccacc
     2221 ctgtactggc acgtgaagaa caagcgcgcc ctgctggacg ccctggccat cgagatgctg
     2281 gaccgccacc acacccactt ctgccccctg gagggcgagt cctggcagga cttcctgcgc
     2341 aacaacgcca agtccttccg ctgcgcactg ctgtcccacc gcgacggcgc caaggtgcac
     2401 ctgggcaccc gccccaccga gaagcagtac gagaccctgg agaaccagct ggccttcctg
     2461 tgccagcagg gcttctccct ggagaacgcc ctgtacgccc tgtccgccgt gggccacttc
     2521 accctgggct gcgtgctgga ggaccaggag caccaggtgg ccaaggagga gcgcgagacc
     2581 cccaccaccg actccatgcc ccccctgctg cgccaggcca tcgagctgtt cgaccaccag
     2641 ggcgccgagc ccgccttcct gttcggcctg gagctgatca tctgcggcct ggagaagcag
     2701 ctgaagtgcg agtccggctc ccccaagaag aagcgcaagg tcgactacaa ggacgacgac
     2761 gacaagccgg ccgacgccct ggacgacttc gacctggaca tgctgccggc cgacgccctg
     2821 gacgacttcg acctggacat gctgccggcc gacgccctgg acgacttcga cctggacatg
     2881 ctgtgataag gatctcgact atccgtacga cgtaccagac tacgcactcg actaagaatt
     2941 atccatccat cacactggcg gccggccgct cgagcatgca tctagagggc cctattctat
     3001 agtgtcacct aaatgctaga ggatctttgt gaaggaacct tacttctgtg gtgtgacata
     3061 attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat ttttaagtgt
     3121 ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa cctatggaac
     3181 tgatgagcgg ccgcaagctt taacagctta tcgataatca acctctggat tacaaaattt
     3241 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3301 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3361 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3421 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3481 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3541 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3601 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3661 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3721 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3781 tctccctttg ggccgcctcc ccgcatcgat accgtcgact cgctgatcag cctcgactgt
     3841 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3901 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3961 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4021 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     4081 cagctggggc tcgactagag catggctacg tagataagta gcatggcggg ttaatcatta
     4141 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4201 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg
     4261 cgcagagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
     4321 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
     4381 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
     4441 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     4501 ttctgcctgc tggggagcct ggggactttc tacacctggt tgctgactaa ttgagatgca
     4561 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     4621 cattccacag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4681 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4741 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4801 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4861 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4921 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4981 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5041 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5101 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5161 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5221 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5281 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5341 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5401 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5461 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5521 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5581 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5641 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5701 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5761 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5821 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5881 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5941 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6001 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6061 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6121 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6181 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6241 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6301 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6361 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6421 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6481 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6541 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6601 ccgaaaagtg ccacctgacg tctaagaaaa cattattatc atgacattaa cctataaaaa
     6661 taggcgtatc acgaggccct ttcgt
//
