LOCUS Exported 7572 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Construct used to synthesize progerin mRNA. Please note the plasmid contains LMNA gene with nucleotide C1824T mutation.. ACCESSION . VERSION . KEYWORDS pAAV-AcGFP-progerin SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7572) AUTHORS Hewitt TITLE pAAV-AcGFP-progerin JOURNAL Unpublished REFERENCE 2 (bases 1 to 7572) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7572 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 60..363 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 364..567 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 518..538 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 564..588 /label=LNCX /note="Human CMV promoter, forward primer" primer_bind 1183..1202 /label=T7 /note="T7 promoter, forward primer" promoter 1183..1201 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 1242..1251 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1248..1964 /codon_start=1 /product="Aequorea coerulescens GFP" /label=AcGFP1 /note="mammalian codon-optimized" /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF GFVTAAAITHGMDELYK" polyA_signal 4173..4649 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4372..4391) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4689..4829 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4689..4818 /label=AAV2 ITR rep_origin 4904..5359 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4991..5010) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5201..5222 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5376..5395) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5495..5517 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5555..5573) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5641..5745 /gene="bla" /label=AmpR promoter CDS 5746..6606 /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(5964..5983) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6777..7365 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7266..7285 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 7427..7556 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" ORIGIN 1 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc 61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg 121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa 181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca 241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac 301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc 361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga 421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg 481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta 541 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc 601 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccgcggattc 661 gaatcccggc cgggaacggt gcattggaac gcggattccc cgtgccaaga gtgacgtaag 721 taccgcctat agagtctata ggcccacaaa aaatgctttc ttcttttaat atactttttt 781 gtttatctta tttctaatac tttccctaat ctctttcttt cagggcaata atgatacaat 841 gtatcatgcc tctttgcacc attctaaaga ataacagtga taatttctgg gttaaggcaa 901 tagcaatatt tctgcatata aatatttctg catataaatt gtaactgatg taagaggttt 961 catattgcta atagcagcta caatccagct accattctgc ttttatttta tggttgggat 1021 aaggctggat tattctgagt ccaagctagg cccttttgct aatcatgttc atacctctta 1081 tcttcctccc acagctcctg ggcaacgtgc tggtctgtgt gctggcccat cactttggca 1141 aagaattggg attcgaacat cgattgaatt ccccggggat cctaatacga ctcactatag 1201 gggaaataag agagaaaaga agagtaagaa gaaatataag agccaccatg gtgagcaagg 1261 gcgccgagct gttcaccggc atcgtgccca tcctgatcga gctgaatggc gatgtgaatg 1321 gccacaagtt cagcgtgagc ggcgagggcg agggcgatgc cacctacggc aagctgaccc 1381 tgaagttcat ctgcaccacc ggcaagctgc ctgtgccctg gcccaccctg gtgaccaccc 1441 tgagctacgg cgtgcagtgc ttctcacgct accccgatca catgaagcag cacgacttct 1501 tcaagagcgc catgcctgag ggctacatcc aggagcgcac catcttcttc gaggatgacg 1561 gcaactacaa gtcgcgcgcc gaggtgaagt tcgagggcga taccctggtg aatcgcatcg 1621 agctgaccgg caccgatttc aaggaggatg gcaacatcct gggcaataag atggagtaca 1681 actacaacgc ccacaatgtg tacatcatga ccgacaaggc caagaatggc atcaaggtga 1741 acttcaagat ccgccacaac atcgaggatg gcagcgtgca gctggccgac cactaccagc 1801 agaatacccc catcggcgat ggccctgtgc tgctgcccga taaccactac ctgtccaccc 1861 agagcgccct gtccaaggac cccaacgaga agcgcgatca catgatctac ttcggcttcg 1921 tgaccgccgc cgccatcacc cacggcatgg atgagctgta caagtccgga cttaaggcct 1981 ctgtcgacag cagtctctgt ccttcgaccc gagccccgcg ccctttccgg gacccctgcc 2041 ccgcgggcag cgctgccaac ctgccggcca tggagacccc gtcccagcgg cgcgccaccc 2101 gcagcggggc gcaggccagc tccactccgc tgtcgcccac ccgcatcacc cggctgcagg 2161 agaaggagga cctgcaggag ctcaatgatc gcttggcggt ctacatcgac cgtgtgcgct 2221 cgctggaaac ggagaacgca gggctgcgcc ttcgcatcac cgagtctgaa gaggtggtca 2281 gccgcgaggt gtccggcatc aaggccgcct acgaggccga gctcggggat gcccgcaaga 2341 cccttgactc agtagccaag gagcgcgccc gcctgcagct ggagctgagc aaagtgcgtg 2401 aggagtttaa ggagctgaaa gcgcgcaata ccaagaagga gggtgacctg atagctgctc 2461 aggctcggct gaaggacctg gaggctctgc tgaactccaa ggaggccgca ctgagcactg 2521 ctctcagtga gaagcgcacg ctggagggcg agctgcatga tctgcggggc caggtggcca 2581 agcttgaggc agccctaggt gaggccaaga agcaacttca ggatgagatg ctgcggcggg 2641 tggatgctga gaacaggctg cagaccatga aggaggaact ggacttccag aagaacatct 2701 acagtgagga gctgcgtgag accaagcgcc gtcatgagac ccgactggtg gagattgaca 2761 atgggaagca gcgtgagttt gagagccggc tggcggatgc gctgcaggaa ctgcgggccc 2821 agcatgagga ccaggtggag cagtataaga aggagctgga gaagacttat tctgccaagc 2881 tggacaatgc caggcagtct gctgagagga acagcaacct ggtgggggct gcccacgagg 2941 agctgcagca gtcgcgcatc cgcatcgaca gcctctctgc ccagctcagc cagctccaga 3001 agcagctggc agccaaggag gcgaagcttc gagacctgga ggactcactg gcccgtgagc 3061 gggacaccag ccggcggctg ctggcggaaa aggagcggga gatggccgag atgcgggcaa 3121 ggatgcagca gcagctggac gagtaccagg agcttctgga catcaagctg gccctggaca 3181 tggagatcca cgcctaccgc aagctcttgg agggcgagga ggagaggcta cgcctgtccc 3241 ccagccctac ctcgcagcgc agccgtggcc gtgcttcctc tcactcatcc cagacacagg 3301 gtgggggcag cgtcaccaaa aagcgcaaac tggagtccac tgagagccgc agcagcttct 3361 cacagcacgc acgcactagc gggcgcgtgg ccgtggagga ggtggatgag gagggcaagt 3421 ttgtccggct gcgcaacaag tccaatgagg accagtccat gggcaattgg cagatcaagc 3481 gccagaatgg agatgatccc ttgctgactt accggttccc accaaagttc accctgaagg 3541 ctgggcaggt ggtgacgatc tgggctgcag gagctggggc cacccacagc ccccctaccg 3601 acctggtgtg gaaggcacag aacacctggg gctgcgggaa cagcctgcgt acggctctca 3661 tcaactccac tggggaagaa gtggccatgc gcaagctggt gcgctcagtg actgtggttg 3721 aggacgacga ggatgaggat ggagatgacc tgctccatca ccaccacggc tcccactgca 3781 gcagctcggg ggaccccgct gagtacaacc tgcgctcgcg caccgtgctg tgcgggacct 3841 gcgggcagcc tgccgacaag gcatctgcca gcggctcagg agcccaggtg ggtggaccca 3901 tctcctctgg ctcttctgcc tccagtgtca cggtcactcg cagctaccgc agtgtggggg 3961 gcagtggggg tggcagcttc ggggacaatc tggtcacccg ctcctacctc ctgggcaact 4021 ccagcccccg aacccagagc ccccagaact gcagcatcat gtaagctgcc ttctgcgggg 4081 cttgccttct ggccatgccc ttcttctctc ccttgcacct gtacctcttg gtctttgaat 4141 aaagcctgag taggaagtct cgagagatct acgggtggca tccctgtgac ccctccccag 4201 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat 4261 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg 4321 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg 4381 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc 4441 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc 4501 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct 4561 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg 4621 cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga 4681 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct 4741 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt 4801 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca 4861 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc 4921 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 4981 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 5041 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 5101 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 5161 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 5221 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt 5281 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa 5341 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag 5401 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc 5461 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt 5521 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag 5581 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg 5641 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 5701 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat 5761 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca 5821 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc 5881 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca 5941 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg 6001 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca 6061 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata 6121 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag 6181 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg 6241 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca 6301 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta 6361 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct 6421 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca 6481 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag 6541 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat 6601 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt 6661 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 6721 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 6781 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 6841 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 6901 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 6961 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 7021 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 7081 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 7141 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 7201 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 7261 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 7321 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 7381 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg 7441 cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc 7501 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg 7561 ccgcacgcgt ct //