LOCUS Exported 7105 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION ssAAV genome with Ple260 (PAX6 MiniPromoter) driving an emerald GFP (EmGFP) reporter. Contains WPRE. ACCESSION . VERSION . KEYWORDS pEMS2049 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7105) 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 7105) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7105 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR intron 2382..2514 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 2497..2516 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 2535..2544 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2541..3260 /codon_start=1 /product="Emerald GFP" /label=EmGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2586..2607) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2847..2866) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3194..3215 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(3326..3346) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3744..3755) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3876..3997 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3887..3906) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3941..3960 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 4166..4295 /label=AAV2 ITR primer_bind complement(4314..4331) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4314..4330) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4323..4345) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4472..4927 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4559..4578) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4769..4790 /label=F1ori-F /note="F1 origin, forward primer" promoter 4953..5057 /gene="bla" /label=AmpR promoter CDS 5058..5918 /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(5276..5295) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6089..6677 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6578..6597 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6831..6848 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6965..6986 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7001..7031 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7039..7055 /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 7044..7066 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7063..7079 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7063..7079 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct 181 aggaagatcg gaattcccta ggccggccac gcgtcagact gggaattgcc gtggctatgc 241 ccgaagtttg gaaagtctct ctcggtctct ttctgggaca gcctttcttt ttccgggaag 301 cttcctgtgc ctagaggtgg ggctctgtag ctggaggggt agatggaact tgcgaagcaa 361 aggaaggcaa taaaaggaag ttaggagaaa ccggaaacgt gggccaggcc ttgctctgtt 421 ggagaagcgg tgtccccgca aagtccctgc ttgcgctggt cccggccgga gcccagatcc 481 cccaaacgcc tgtcacttct catcgagttg aggctccggc ccccacccgc gccttggtga 541 gggccccccc tcagccccaa gccttcggct accctgaaaa cgcggctccc ctcgaagggg 601 aaacaagtgc agttcaatct cgtctgagtg atctacaaat aaggacggaa agggtcgttt 661 tatcacggtc ccgtttgtcg tgacgatgca atttcccgga gcggagcact gtcacaaagt 721 gacaaggctg ccacaagcgc cccgactgat cttttcaatt agccttccat gcatgatccg 781 gagcgacttc cgcctatttc cagaaattaa gctcaaactt gacgtgcagc tagttttatt 841 ttaaagacaa atgtcagaga ggctcatcat attttccccc ctcttctata tttggagctt 901 atttattgct aagaagctca ggctcctggc gtcaatttat cagtaggctc caaggagaag 961 agaggagagg agaggagagc tgaacaggga gccacgtctt ttcctgggag ggctgctatc 1021 taagtcgggg ctgcaggttg gagattttta aggaagtgga aattggcaat tggctttgtg 1081 tgtctgtggt ttttggggag ggggactaca aagaggggct aactccctct ccctattctc 1141 taaggttgga ccacagggat gaggttgtga gatacaaaga taaaggaggg atggggaaca 1201 ctatgatgtg gtatttttct tttctgtttt ttctttttga taatatctat ccttggctaa 1261 ggggagggcg gagttcagcg gcggaaataa agcgagcagt ggctggtgcg aaccgactcc 1321 cggctctaag ccttacttgc ggtggccgga cttgtctgtg gctgaagccg agcccgggct 1381 ctgactctca cgtctgcact ggaggtgcgg aaacctggac tggggttcac cagccacata 1441 ctggctgctc tggttgttct ctcctcttcc ttcttctatt ctaaccaaac aaaacccaac 1501 tcgatgcttg tgccaggcct tggccagttt ggacggtgat ggaaacattt ctggatttta 1561 gcgtctaagg gagcactcaa gggctgtaag gtttgctatc actgtcccaa cactgcagag 1621 accttgaagg ttcagtgtgg gatctgtaga acccatggtt tgcggtgaca ctcagagggg 1681 atctttggga gatttataga gccggttctt agcgctttgt gggttcagag gctggctgca 1741 gtgtttatga agaggggcag tgggctgggg acactgctgg ggttatggct gtagtgaggt 1801 ccatgtgtac ttgttccttg gcatgtgtct gactctgtgt tgctgctgca gtacagtggg 1861 caggggcacg gttgcttgga ctgggctgcc cgatcaccct ccgaggactg ggagtcccga 1921 gtccagctta gggggagtgg gggcgcgacc cccaacccag aaaccttcac ttgaccgctc 1981 aagttcgcgg cagcagggcg ggccgcgccg aatctcggcg tgcgcggagc ggggagatgc 2041 aggcgagcgc cagagcccgg gctcgggggc cctgcgccgg ggagaggagc cgggacccac 2101 cggcggagcc gaaaacaagt gtattcatat tcaaacaaac ggaccaattg caccaggcgg 2161 ggagagggag catccaatcg gctggcgcga ggccccggcg ctgctttgca taaagcaata 2221 ttttgtgtga gagcgagcgg tgcatttgca tgttgcggag tgattagtgg gtttgaaaag 2281 ggaaccgtgg ctcggcctca tttcccgctc tggttcaggc gcaggaggaa gtgttttgct 2341 ggaggatggc gcgccgaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa 2401 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt 2461 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc 2521 actcccaggc ggccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc 2581 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg 2641 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc 2701 tgcccgtgcc ctggcccacc ctcgtgacca ccttcaccta cggcgtgcag tgcttcgccc 2761 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg 2821 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga 2881 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg 2941 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaag gtctatatca 3001 ccgccgacaa gcagaagaac ggcatcaagg tgaacttcaa gacccgccac aacatcgagg 3061 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg 3121 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg 3181 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca 3241 tggacgagct gtacaagtaa gcggccgcga tcgctcaacc tctggattac aaaatttgtg 3301 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 3361 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 3421 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 3481 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 3541 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 3601 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 3661 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg 3721 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 3781 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 3841 ccctttgggc cgcctccccg cgatcgcaga catgataaga tacattgatg agtttggaca 3901 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc 3961 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt 4021 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa 4081 atgtggtagt cgaaattccc gataaggatc ttcctagagc atggctacgt agataagtag 4141 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct 4201 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt 4261 gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta attaacctaa ttcactggcc 4321 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca 4381 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc 4441 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg 4501 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 4561 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 4621 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 4681 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 4741 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 4801 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 4861 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt 4921 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 4981 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 5041 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 5101 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 5161 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 5221 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 5281 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact 5341 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 5401 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 5461 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 5521 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 5581 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg 5641 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 5701 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 5761 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 5821 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 5881 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 5941 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 6001 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 6061 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 6121 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 6181 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc 6241 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 6301 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 6361 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 6421 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 6481 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 6541 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 6601 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 6661 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 6721 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 6781 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag 6841 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga 6901 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg 6961 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg 7021 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc 7081 atgattacgc cagatttaat taagg //