LOCUS Exported 6312 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cre-activated AAV expression of mCherry. ACCESSION . VERSION . KEYWORDS pAAV-EF1a-double floxed-mCherry-WPRE-HGHpA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6312) TITLE Cre-activated AAV expression plasmids REFERENCE 2 (bases 1 to 6312) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6312 /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)." CDS complement(1576..2286) /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1854..1873) /label=mCherry-F /note="mCherry, forward primer" primer_bind 2123..2141 /label=mCherry-R /note="mCherry, reverse primer" regulatory 2283..2292 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(2297..2330) /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(2381..2414) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 2439..3027 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2492..2512) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2910..2921) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3059..3535 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3258..3277) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3575..3715 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3575..3704 /label=AAV2 ITR rep_origin 3790..4245 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3877..3896) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4087..4108 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4262..4281) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4381..4403 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4441..4459) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4527..4631 /gene="bla" /label=AmpR promoter CDS 4632..5492 /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(4850..4869) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5663..6251 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6152..6171 /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 gcgccttact tgtacagctc gtccatgccg ccggtggagt ggcggccctc 1621 ggcgcgttcg tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt 1681 gatgttgacg ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt 1741 gacctcagcg tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt 1801 cagggcgccg tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt 1861 ctgcattacg gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa 1921 ctcgccgtcc tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt 1981 catcacgcgc tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc 2041 ggcggggtgc ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca 2101 ggcgaagggc agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta 2161 ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat 2221 gtgcaccttg aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct 2281 caccatggtg gctagcataa cttcgtataa agtatcctat acgaagttat ttgccttaac 2341 ccagaaatta tcactgttat tctttagaat ggtgcaaaga ataacttcgt ataatgtatg 2401 ctatacgaag ttatgaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa 2461 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 2521 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 2581 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 2641 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 2701 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 2761 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 2821 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc 2881 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 2941 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 3001 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg 3061 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt 3121 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt 3181 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac 3241 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct 3301 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt 3361 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg 3421 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg 3481 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat 3541 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc 3601 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc 3661 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc 3721 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc 3781 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 3841 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 3901 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt 3961 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac 4021 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 4081 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt 4141 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac 4201 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca 4261 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg 4321 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct 4381 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg 4441 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt 4501 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 4561 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 4621 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 4681 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 4741 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 4801 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 4861 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 4921 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 4981 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 5041 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 5101 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 5161 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 5221 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 5281 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 5341 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 5401 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 5461 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 5521 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 5581 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 5641 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 5701 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 5761 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 5821 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 5881 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 5941 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 6001 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 6061 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 6121 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 6181 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 6241 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 6301 ttgctcacat gt //