LOCUS Exported 6685 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses TetActivator protein under the control of the c-fos promoter for use in TetTaging. ACCESSION . VERSION . KEYWORDS pAAV-cFos-tTA-pA 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6685 /organism="synthetic DNA construct" /mol_type="other DNA" 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 complement(4339..4358) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 4606..4626 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" primer_bind complement(4696..4713) /label=L4440 /note="L4440 vector, forward primer" 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(5626..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 //