LOCUS Exported 7673 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION ssAAV genome with Ple343 (TUBB3 MiniPromoter) driving an emerald GFP (EmGFP) reporter. Contains WPRE. ACCESSION . VERSION . KEYWORDS pEMS2273 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7673) AUTHORS Simpson E, Korecki AJ, Fornes O, McGill TJ, Cueva-Vargas JL, Agostinone J, Farkas RA, Hickmott JW, Lam SL, Mathelier A, Renner LM, Stoddard J, Zhou M, Di Polo A, Neuringer M, Wasserman WW TITLE New MiniPromoter Ple345 (NEFL) Drives Strong and Specific Expression in Retinal Ganglion Cells of Mouse and Primate Retina. JOURNAL Hum Gene Ther. 2018 Jul 31. doi: 10.1089/hum.2018.118. PUBMED 30062914 REFERENCE 2 (bases 1 to 7673) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7673 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR intron 2912..3044 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 3027..3046 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 3065..3074 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3071..3790 /codon_start=1 /product="Emerald GFP" /label=EmGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3116..3137) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3377..3396) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3724..3745 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(3856..3876) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4274..4285) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4406..4527 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4417..4436) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4471..4490 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 4723..4852 /label=AAV2 ITR primer_bind complement(4871..4888) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4871..4887) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4880..4902) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 5029..5484 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5116..5135) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5326..5347 /label=F1ori-F /note="F1 origin, forward primer" promoter 5510..5614 /gene="bla" /label=AmpR promoter CDS 5615..6475 /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(5833..5852) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6646..7234 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7135..7154 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7388..7405 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7522..7543 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7558..7588 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7596..7612 /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 7601..7623 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7620..7636 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7620..7636 /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 ggccggcccc tgcgctggga gccgagcccc cggcagctcg 241 cggggatgca gtaagccggg cgcagcccgc cgggaggatg agaaaacccg aagcccggaa 301 agccgagcgg ccgcctgcag tcaccccggg gggggcaggg gcgggccggg ctatgcagaa 361 acaccggggc ccgcgggaca caggacgctg ccccactgcc gcagagctga atctggaccc 421 ccgggctttg ctttcctggg gcctgtgctc ctccctcccc ggggcggggt tcgcctgcag 481 cggcggaggg gggagccttt gtcctccgcg cgggctgcgg caccgcctcc tcgcggctgc 541 ggggcggggc tgggggcggg gcctggccgt ctcagccaat gggaggcgga gtccctggct 601 cgccccgccc tctcgcctga aagacctgcg gagccgcggg ccgggcggga accgcattcg 661 ggtcctggag gggcttggag gccccagtct cgcagcccct tcatcgggga cccccgctcc 721 cacctgcctt ggccactgcc ctcaaagccc ggccccgctt tccccaggac cccaagcgct 781 gcgcgaggtc agcaccaagg acagcgcccg ggccgagcgc ctggcctcga acgccgggtt 841 ctgctttgtg tttccaggtt tttgtttttg gttatgagag gaggcctggc tctgtcgccc 901 gggctggcgt ccagaggcgc gatctcggct cactgcagcc tcggcctccc gagtagctgg 961 gaccgcaggc gctcaccacc acgcctggcc acaattttcc ttttttctta ttttctttta 1021 gatggagttt ggctcttgtt gcccaggctg ggggacgtcg tgtcccctgg tcctgggagg 1081 ctgtccctgg cccaaggtca cacagcaggt gcgagagagc tgggctggcc cttggttctc 1141 atcccctgca ggagccaacc tggggctcgg gcggggcggg ttgggggcgc tgcttgccaa 1201 aggatctgcc ctccgaacag tgttggggaa caggggatcg gggggggagg ctggctttgt 1261 caggaccaag gccagggacc cagctcaggg ctggtgcagc gctgattctg ggaaggaggg 1321 ggtgcaggag acggtcctgg aggggaaggg gccagagaaa ggcagcctcc ggaggctgtt 1381 gccatggaaa ccaggcagga acaaaaagct aattacaaca gggccagcca cgtggctgac 1441 atgtaaattg caactttggc tccgggtgga ggcgcctctg agggtgggag gcccagtagt 1501 ggggagactc taatcaccca aggagacagc tgtgcctgcc aaggagagcc tgggggaggg 1561 gagacctggg ggacagtggc ctcaaatcaa accaggactc ccggagcagc ctctgacagc 1621 tgccttggcc tgatgggagg ggctgagggc tcattaatgt ggtcactggg gccaagcctc 1681 ccctgagcat ctgctcctgg gagggtcagg ggtcgctggg gggagggtcg cagctgctca 1741 ttttgtgaag tgaatgctga tttgggggcc gggtggggtt cgtctgtaca tcgtgcactg 1801 tcagaccctt cctgaaggat ttttgttact gaagtatcag aaggcccttg ttctaaggtg 1861 gtggcactgt aatatctggc ccaggaggag gtcagtgccg tgccggcgag tcctttgctc 1921 ccttccttct ctgaccagta aacgcatctc cagtcgctcc cccgaggccc ccctgcacgc 1981 agtctccgac ctcagagagt gagtcagata gaagccgctg cccaccctcc cccaccctgg 2041 ccgggttctt gcctggcttg tgcgtccttg tgacaattcc tgacacggat gcgcaccggg 2101 cagtggcacg tccaggtgct gtgcgcgccc gggcccacaa agctcctttg gacgctcatg 2161 gcagcccctt ggtggaggag gtcgggtgcc caccccctcc ccgcccactg cggaagccgg 2221 cgacccacgg agctcgctct cggccgccgc cacccctctg tgttcgcgcc cttccgagct 2281 ctgatccgac gctttgtttc ttctcagtgg gttcagggcc tgggccagcc tttacctacc 2341 tcccccaccc aaaaccggca aaagctcaga gcaccttgtc tgccaaaaga cagggagctg 2401 ggatggtgcg ggttggtctc taaaccggcg tggggaaaaa agaccctccg tacaaagccg 2461 cagggtgggg ctgtcgcaag ggcggaaccg agagggtagc tgggggcggg gttcccaggg 2521 ccaagagggg ccattgtcct ccctggagcc cggcgccccc acagccagct cctctgggag 2581 acagcccctc ctttcgaatg cgcggggccc tcagaccgcg cccggcccag cgctggggga 2641 tccttggctg cgggaggggc gccgcattgc gcgcggcggg cggggacgcg cggtgcggag 2701 cctgcgggcc gggcggggct ctgcggcggc gcctcccgat tggccacccg cggtgacatc 2761 agccgatgcg aagggcgggg ccgcggctat aagagcgcgc ggccgcggtc cccgaccctc 2821 agcagccagc ccggcccgcc cgcgcccgtc cgcagccgcc cgccagacgc gcccagtggc 2881 gcgccgaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta 2941 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc 3001 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccaggc 3061 ggccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt 3121 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga 3181 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc 3241 ctggcccacc ctcgtgacca ccttcaccta cggcgtgcag tgcttcgccc gctaccccga 3301 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg 3361 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg 3421 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat 3481 cctggggcac aagctggagt acaactacaa cagccacaag gtctatatca ccgccgacaa 3541 gcagaagaac ggcatcaagg tgaacttcaa gacccgccac aacatcgagg acggcagcgt 3601 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc 3661 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga 3721 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct 3781 gtacaagtaa gcggccgcga tcgctcaacc tctggattac aaaatttgtg aaagattgac 3841 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 3901 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 3961 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 4021 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 4081 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 4141 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc 4201 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt 4261 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 4321 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc 4381 cgcctccccg cgatcgcaga catgataaga tacattgatg agtttggaca aaccacaact 4441 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta 4501 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag 4561 gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtagt 4621 cgaaattccc gataaggatc ttcgtcgacg aattcccgat aaggatcttc ctagagcatg 4681 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg 4741 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg 4801 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agccttaatt 4861 aacctaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc 4921 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc 4981 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta 5041 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca 5101 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct 5161 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc 5221 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat 5281 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc 5341 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc 5401 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta 5461 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc 5521 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg 5581 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc 5641 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt 5701 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct 5761 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac 5821 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact 5881 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa 5941 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga 6001 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt 6061 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga 6121 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg 6181 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat 6241 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat 6301 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc 6361 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga 6421 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc 6481 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag 6541 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc 6601 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt 6661 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt 6721 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat 6781 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 6841 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 6901 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 6961 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 7021 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 7081 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa 7141 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt 7201 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 7261 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc 7321 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac 7381 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct 7441 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc 7501 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt 7561 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac 7621 aggaaacagc tatgaccatg attacgccag atttaattaa ggccttaatt agg //