LOCUS Exported 5180 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION An AAV genome with tet-inducible expression of the fluorescent protein eYFP. ACCESSION . VERSION . KEYWORDS pAAV-TRE-eYFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5180) AUTHORS Challis RC, Ravindra Kumar S, Chan KY, Challis C, Beadle K, Jang MJ, Kim HM, Rajendran PS, Tompkins JD, Shivkumar K, Deverman BE, Gradinaru V TITLE Systemic AAV vectors for widespread and targeted gene delivery in rodents. JOURNAL Nat Protoc. 2019 Feb;14(2):379-414. doi: 10.1038/s41596-018-0097-3. PUBMED 30626963 REFERENCE 2 (bases 1 to 5180) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5180 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" protein_bind 165..183 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 202..220 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 239..257 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 276..294 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 313..331 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 350..368 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 387..405 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 424..442 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 464..488 /label=LNCX /note="Human CMV promoter, forward primer" CDS 563..1282 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(608..629) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(869..888) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1216..1237 /label=EGFP-C /note="EGFP, forward primer" misc_feature 1307..1895 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1360..1380) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1778..1789) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 1927..2403 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2126..2145) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 2443..2583 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2443..2572 /label=AAV2 ITR rep_origin 2658..3113 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2745..2764) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2955..2976 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3130..3149) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3249..3271 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3309..3327) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3395..3499 /gene="bla" /label=AmpR promoter CDS 3500..4360 /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(3718..3737) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4531..5119 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5020..5039 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtatcaag tgccacctga cgtctcccta tcagtgatag 181 agaagtcgac acgtctcgag ctccctatca gtgatagaga aggtacgtct agaacgtctc 241 cctatcagtg atagagaagt cgacacgtct cgagctccct atcagtgata gagaaggtac 301 gtctagaacg tctccctatc agtgatagag aagtcgacac gtctcgagct ccctatcagt 361 gatagagaag gtacgtctag aacgtctccc tatcagtgat agagaagtcg acacgtctcg 421 agctccctat cagtgataga gaagctaccc cctatataag cagagctcgt ttagtgaacc 481 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc 541 gatccagcct ggatccggta ccatggtgag caagggcgag gagctgttca ccggggtggt 601 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 661 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 721 gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc 781 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 841 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 901 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 961 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 1021 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 1081 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 1141 cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa 1201 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 1261 catggacgag ctgtacaagt aagaattcga tatcaagctt atcgataatc aacctctgga 1321 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg 1381 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt 1441 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag 1501 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc 1561 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga 1621 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa 1681 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac 1741 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct 1801 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca 1861 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag 1921 atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc 1981 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag 2041 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg 2101 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa 2161 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc 2221 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag 2281 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac 2341 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc 2401 cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg 2461 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg 2521 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc 2581 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac 2641 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 2701 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt 2761 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 2821 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga 2881 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 2941 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg 3001 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct 3061 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg 3121 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac 3181 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt 3241 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag 3301 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc 3361 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 3421 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 3481 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt 3541 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc 3601 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat 3661 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct 3721 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca 3781 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg 3841 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa 3901 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg 3961 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga 4021 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg 4081 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt 4141 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg 4201 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc 4261 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca 4321 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc 4381 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 4441 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 4501 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 4561 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 4621 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 4681 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 4741 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 4801 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 4861 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 4921 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 4981 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 5041 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 5101 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 5161 ctggcctttt gctcacatgt //