LOCUS Exported 5841 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV-mediated expression of tdTomato under the TRE promoter.. ACCESSION . VERSION . KEYWORDS pAAV-TRE-tdTomato-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5841) 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 5841) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5841 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 94..111 /label=L4440 /note="L4440 vector, forward primer" protein_bind 228..249 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 264..294 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 302..318 /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)." primer_bind 307..329 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 326..342 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 326..342 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" repeat_region 380..509 /label=AAV2 ITR protein_bind 583..601 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 619..637 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 654..672 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 690..708 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 726..744 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 761..779 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 797..815 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" promoter 827..865 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 862..886 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 900..909 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 906..2336 /codon_start=1 /product="tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)" /label=tdTomato /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE RSEGRHHLFLYGMDELYK" misc_feature 2350..2938 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2403..2423) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2821..2832) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2941..2957) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2942..2958) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 2987..3108 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2998..3017) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3052..3071 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3270..3399 /label=AAV2 ITR primer_bind complement(3418..3435) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3418..3434) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3427..3449) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3576..4031 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3663..3682) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3873..3894 /label=F1ori-F /note="F1 origin, forward primer" promoter 4057..4161 /gene="bla" /label=AmpR promoter CDS 4162..5022 /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(4380..4399) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5193..5781 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5682..5701 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag 241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga 301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccagattt 361 aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc 421 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg 481 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt 541 atctacgtag ccatgctcta ggaagatctc gaattggttt actccctatc agtgatagag 601 aacgtatgtc gagtttactc cctatcagtg atagagaacg atgtcgagtt tactccctat 661 cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag 721 tttactccct atcagtgata gagaacgtat gtcgagttta tccctatcag tgatagagaa 781 cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgaggta ggcgtgtacg 841 gtgggaggcc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagaattcg 901 ccaccatggt gagcaagggc gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca 961 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct 1021 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct 1081 gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg 1141 acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga 1201 acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc 1261 tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga 1321 agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga 1381 agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca 1441 agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca 1501 agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg 1561 agggccgcca ccacctgttc ctggggcatg gcaccggcag caccggcagc ggcagctccg 1621 gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa agagttcatg cgcttcaagg 1681 tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc 1741 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct 1801 tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac gtgaagcacc 1861 ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag tgggagcgcg 1921 tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg 1981 gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac ggccccgtaa 2041 tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg 2101 tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac tacctggtgg 2161 agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac tactacgtgg 2221 acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc 2281 gctccgaggg ccgccaccac ctgttcctgt acggcatgga cgagctgtac aagtaagaag 2341 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 2401 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 2461 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 2521 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 2581 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 2641 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 2701 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct 2761 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 2821 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 2881 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat 2941 cgataccgtc gacctcgacc cgggcggccg cttcgagcag acatgataag atacattgat 3001 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt 3061 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat 3121 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa 3181 aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa 3241 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc 3301 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg 3361 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc cttaattaac ctaattcact 3421 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct 3481 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc 3541 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag 3601 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 3661 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 3721 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 3781 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 3841 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 3901 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 3961 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac 4021 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt 4081 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa 4141 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt 4201 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat 4261 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag 4321 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg 4381 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata 4441 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat 4501 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc 4561 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg 4621 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac 4681 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact 4741 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa 4801 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct 4861 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc 4921 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga 4981 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac 5041 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag 5101 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg 5161 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc 5221 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag 5281 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt 5341 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac 5401 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc 5461 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt 5521 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt 5581 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc 5641 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt 5701 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca 5761 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt 5821 tgctggcctt ttgctcacat g //