LOCUS Exported 7129 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION ssAAV genome with Ple257 (PAX6 MiniPromoter) driving an emerald GFP (EmGFP) reporter. Contains WPRE. ACCESSION . VERSION . KEYWORDS pEMS2046 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7129) AUTHORS Hickmott JW, Chen CY, Arenillas DJ, Korecki AJ, Lam SL, Molday LL, Bonaguro RJ, Zhou M, Chou AY, Mathelier A, Boye SL, Hauswirth WW, Molday RS, Wasserman WW, Simpson EM TITLE PAX6 MiniPromoters drive restricted expression from rAAV in the adult mouse retina. JOURNAL Mol Ther Methods Clin Dev. 2016 Aug 10;3:16051. doi: 10.1038/mtm.2016.51. eCollection 2016. PUBMED 27556059 REFERENCE 2 (bases 1 to 7129) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7129 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 12..141 /label=AAV2 ITR intron 2406..2538 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 2521..2540 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 2559..2568 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2565..3284 /codon_start=1 /product="Emerald GFP" /label=EmGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2610..2631) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2871..2890) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3218..3239 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(3350..3370) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3768..3779) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3900..4021 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3911..3930) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3965..3984 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 4190..4319 /label=AAV2 ITR primer_bind complement(4338..4355) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4338..4354) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4347..4369) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4496..4951 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4583..4602) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4793..4814 /label=F1ori-F /note="F1 origin, forward primer" promoter 4977..5081 /gene="bla" /label=AmpR promoter CDS 5082..5942 /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(5300..5319) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6113..6701 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6602..6621 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6855..6872 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6989..7010 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7025..7055 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7063..7079 /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 7068..7090 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7087..7103 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7087..7103 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 ccttaattag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt 181 agccatgctc taggaagatc ggaattccct aggccggcca cgcgtctcca ctcctatccc 241 atccccacgt ccccgaaatg aataatgcag gcaggagcca agatttctgc attagcgcca 301 cgttgccccg gcggctggcg ctaggatccc gaagtcggat aatggaaatt aaactgttgt 361 tgtgtggcgg ggatgtcatt atgtgactgt tcatattcaa agtcaggcag caatgaattg 421 taatcatttc aattatcttc aatacacttt ctttaggaca caagttgtca aggataaaaa 481 aaaaaagtac attcttgcgg ctccgtgggt ccattttcca gatggtttgt tactcttgct 541 gcctgattta gtttcggctt ttattcccca gcctcagatt ctcttgcaga gactccgccg 601 gagtgagcgg cttctctggc gctgcgggcc ggctacgctg gtcggcaatc ggcctccgtg 661 ggcctggcgt gcgcattttg gttgctttca ggtataatta actttgaaca acaaatatgc 721 ctggagctct ggatctagtg tccccttggg gaagggggcc gtcgtattat actctgcgtc 781 cttcccagcc tggggcgggg tgggggctgg ctgcagcagc tgcacccact ccccaggcca 841 gcgctgcgcg tcagaaaagc atgtgctggg ctgtacccac tcctccgaca gagcccggac 901 cctctccttc caacccagag aactcaggga aaagccggga tccccggagc ctgccgtcct 961 ttgttgaaat taccctctgc ccaatgttcg ctccctcctg tgcctgaacg tggggctgcg 1021 acaagttgga tgtggtgtcc ttgaagtatc ccttcgagcg ccctccgccc ccagactggg 1081 caaaaagagt aggggtcgcc tttgaagccc aagtccgcgg ctcctatttg gactaatccc 1141 gatctttaca ctaatatcgc ctctcgggtt ttcattatac tcatttggga ggaaatgagc 1201 ttttgctggg caaatagcac cctgcacccc acttctcata gctgtgcagg cccggggcct 1261 gaacaacccc gcaggcaatc cgggagctgc cgatattata tttgtgaaat gtccttttgt 1321 tgaggctttt aacaccttct tataacgaat catttgttct tttctcttta attttgatag 1381 acaattagcc atcagagata caccagttga ctgtaaccga gcattaaact cgcctttatg 1441 taacgctaat ttagcatagc tgctttgttt aactttgcta tataagaacc aggttttgta 1501 gcaaaaaaaa aaaggtgtaa ttttatgtgg ggcttcattt cttttggccc tgtacacagc 1561 tagccctctc ctctttgtga agcgttctgc tggggcctga cgccatttcc cgtgcctagc 1621 acctgcaggc tggggggaga gagggaatta gcattgatat agatttagtt gaaatgtaat 1681 ctcttaaatc tgtccaagat tcggggagat aattacactt tcactgaatt gccaggggtc 1741 agcgtctctg aggacctgag gagttgattt actaacaata gtgttacata aaaaatggcg 1801 tttgagatta taatgttaat ggctgcggtc gctggggaat gcgcccgctt ccaactaatt 1861 ccatgtaact tgtttttttt cctccctttt cctccctccc tttcttccag gctccgtctt 1921 ggggatcgga ggaggggaca gggtgattac ccagagaggt agctggccag cctaagggca 1981 gagatcttgg ggccctagtg cccgaaggtg cggaggagcg cactcggcaa gactagtttc 2041 ctggggatcg actctacgcc atacaggacg gcggcccagg ctggaccggg ccgggctaga 2101 gcagtcacag gccgggccaa ggaaggccaa agcaggggtt ggagccggcc ggaccctggg 2161 tggggagaag caggctcccg cccggccgga aaactagtcg gcgcagagct gtgcccaact 2221 ctagccgcca tgacgtcacg cgggccgggc agccaatgag gacggcgctg gcgtggatat 2281 taaggaaagt tagcgcctgc ctgagcaccc tcttttctta tcattgacat ttaaactctg 2341 gggcaggtcc tcgcgtagaa cgcggctgtc aggcgcgccg aagttggtcg tgaggcactg 2401 ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt 2461 cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact 2521 ttgcctttct ctccacaggt gtccactccc aggcggccgc caccatggtg agcaagggcg 2581 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc 2641 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga 2701 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttca 2761 cctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca 2821 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca 2881 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc 2941 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact 3001 acaacagcca caaggtctat atcaccgccg acaagcagaa gaacggcatc aaggtgaact 3061 tcaagacccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga 3121 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt 3181 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga 3241 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagcggcc gcgatcgctc 3301 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 3361 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 3421 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 3481 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 3541 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 3601 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 3661 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct 3721 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 3781 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 3841 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgatcg cagacatgat 3901 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat 3961 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt 4021 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt 4081 ttaaagcaag taaaacctct acaaatgtgg tagtcgaaat tcccgataag gatcttccta 4141 gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta 4201 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca 4261 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc 4321 cttaattaac ctaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc 4381 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa 4441 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg 4501 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 4561 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 4621 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 4681 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 4741 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 4801 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 4861 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 4921 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg 4981 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 5041 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc 5101 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 5161 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac 5221 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga 5281 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag 5341 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca 5401 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca 5461 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa 5521 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc 5581 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa 5641 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag 5701 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct 5761 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac 5821 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa 5881 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt 5941 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat 6001 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg 6061 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 6121 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 6181 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 6241 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 6301 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 6361 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 6421 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 6481 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 6541 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 6601 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 6661 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 6721 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc 6781 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc 6841 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac 6901 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact 6961 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc 7021 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat 7081 ttcacacagg aaacagctat gaccatgatt acgccagatt taattaagg //