LOCUS Exported 4113 bp ds-DNA circular SYN 16-MAR-2022 DEFINITION AAV vector backbone designed to express GFP from a TRE3G promoter. It also contains a poly-Adenylation Signal (pA). ACCESSION . VERSION . KEYWORDS pJEP205-AAV-TRE3G-GFP-pA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4113) AUTHORS Holehonnur R, Lella SK, Ho A, Luong JA, Ploski JE TITLE The production of viral vectors designed to express large and difficult to express transgenes within neurons. JOURNAL Mol Brain. 2015 Feb 24;8:12. doi: 10.1186/s13041-015-0100-7. PUBMED 25887710 REFERENCE 2 (bases 1 to 4113) AUTHORS . TITLE Direct Submission JOURNAL Exported Mar 16, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4113 /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 175..193 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 211..229 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 247..265 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 283..301 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 319..337 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 355..373 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 391..409 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 454..478 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 555..564 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 561..1277 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(606..627) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(867..886) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1214..1235 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1318..1337) /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 1376..1516 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 1376..1505 /label=AAV2 ITR rep_origin 1591..2046 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1678..1697) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1888..1909 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2063..2082) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2182..2204 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2242..2260) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2328..2432 /gene="bla" /label=AmpR promoter CDS 2433..3293 /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(2651..2670) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3464..4052 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3953..3972 /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 gcgtatcacg agactagcct cgagttggct ttactcccta 181 tcagtgatag agaacgtatg aagagtttac tccctatcag tgatagagaa cgtatgcaga 241 ctttactccc tatcagtgat agagaacgta taaggagttt actccctatc agtgatagag 301 aacgtatgac cagtttactc cctatcagtg atagagaacg tatctacagt ttactcccta 361 tcagtgatag agaacgtata tccagtttac tccctatcag tgatagagaa cgtataagct 421 ttaggcgtgt acggtgggcg cctataaaag cagagctcgt ttagtgaacc gtcagatcgc 481 ctggagcaat tccacaacac ttttgtctta taccaacttt ccgtaccact tcctaccctc 541 gtaaagaatt cgccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc 601 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg 661 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc 721 tgcccgtgcc ctggcccact ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc 781 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg 841 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga 901 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg 961 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca 1021 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg 1081 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg 1141 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg 1201 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca 1261 tggacgagct gtacaagtaa ctcgagttta ttgcagctta taatggttac aaataaagca 1321 atagcatcac aaatttcaca aataaagcat ttttttcact ggaccgagcg gccgcaggaa 1381 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg 1441 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg 1501 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt 1561 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg 1621 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc 1681 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 1741 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 1801 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 1861 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 1921 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt 1981 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta 2041 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc 2101 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt 2161 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac 2221 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga 2281 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta 2341 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 2401 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc 2461 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga 2521 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca 2581 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt 2641 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg 2701 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc 2761 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata 2821 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt 2881 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag 2941 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca 3001 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg 3061 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg 3121 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag 3181 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg 3241 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag 3301 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga 3361 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt 3421 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc 3481 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 3541 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac 3601 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac 3661 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt 3721 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct 3781 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat 3841 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt 3901 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg 3961 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt 4021 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt 4081 tcctggcctt ttgctggcct tttgctcaca tgt //