LOCUS Exported 6470 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pAAV-Ef1a-DO-EGFP-WPRE-pA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6470) AUTHORS Saunders A, Johnson CA, Sabatini BL TITLE Novel recombinant adeno-associated viruses for Cre activated and inactivated transgene expression in neurons. JOURNAL Front Neural Circuits. 2012 Jul 27;6:47. doi: 10.3389/fncir.2012.00047. eCollection 2012. PUBMED 22866029 REFERENCE 2 (bases 1 to 6470) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6470 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR promoter 242..1419 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 473..1410 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1367..1387 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 1450..1483 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 1534..1567 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 1589..1598 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1595..2440 /codon_start=1 /product="EGFP destabilized by fusion to a mutated version of residues 422-461 of mouse ornithine decarboxylase, giving an in vivo half-life of ~1 hour" /label=d1EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYKKLSHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV " primer_bind complement(1640..1661) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1901..1920) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2248..2269 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(2455..2488) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(2539..2572) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 2597..3185 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2650..2670) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3068..3079) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3217..3693 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3416..3435) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3733..3873 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3733..3862 /label=AAV2 ITR rep_origin 3948..4403 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4035..4054) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4245..4266 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4420..4439) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4539..4561 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4599..4617) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4685..4789 /gene="bla" /label=AmpR promoter CDS 4790..5650 /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(5008..5027) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5821..6409 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6310..6329 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt 181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat 241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg 301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag 361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc 421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc 481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat 541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc 1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga 1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg 1561 aagttatggc gcgccttatc tagatttcgc caccatggtg agcaagggcg aggagctgtt 1621 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag 1681 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg 1741 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt 1801 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat 1861 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac 1921 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat 1981 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca 2041 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg 2101 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat 2161 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag 2221 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg 2281 gatcactctc ggcatggacg agctgtacaa gaagcttagc catggcttcc cgccggcggt 2341 ggcggcgcag gatgatggca cgctgcccat gtcttgtgcc caggagagcg ggatggaccg 2401 tcaccctgca gcctgtgctt ctgctaggat caatgtgtag gcggccgcgc tagcataact 2461 tcgtataaag tatcctatac gaagttattt gccttaaccc agaaattatc actgttattc 2521 tttagaatgg tgcaaagaat aacttcgtat aatgtatgct atacgaagtt atgaattcga 2581 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat 2641 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca 2701 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc 2761 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc 2821 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt 2881 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg 2941 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc 3001 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta 3061 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg 3121 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc 3181 cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc 3241 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata 3301 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg 3361 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat 3421 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg 3481 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc 3541 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg 3601 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta 3661 caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga 3721 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct 3781 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct 3841 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta 3901 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag 3961 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 4021 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 4081 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca 4141 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata 4201 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 4261 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc 4321 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa 4381 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc 4441 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 4501 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 4561 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt 4621 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa 4681 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 4741 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca 4801 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca 4861 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta 4921 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt 4981 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc 5041 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc 5101 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc 5161 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa 5221 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga 5281 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat 5341 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca 5401 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc 5461 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat 5521 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag 5581 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa 5641 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca 5701 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 5761 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 5821 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 5881 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 5941 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 6001 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 6061 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 6121 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 6181 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 6241 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 6301 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 6361 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 6421 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt //