LOCUS Exported 3608 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CMV driven Multi Cloning Site-2. ACCESSION . VERSION . KEYWORDS pJEP300-pAAV-CMV-MCS2-pA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3608) AUTHORS Kumar N, Stanford W, Aradhana CS, Abraham ND, Dao TJ, Thaseen S, Sairavi A, Gonzalez CU, Ploski JE. TITLE The Development of an AAV-Based CRISPR SaCas9 Genome Editing System That Can Be Delivered to Neurons in vivo and Regulated via Doxycycline and Cre-Recombinase. JOURNAL Front. Mol. Neurosci. (2018) 13 REFERENCE 2 (bases 1 to 3608) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3608 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" enhancer 154..457 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 458..661 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 612..632 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 658..682 /label=LNCX /note="Human CMV promoter, forward primer" primer_bind complement(813..832) /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 871..1011 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 871..1000 /label=AAV2 ITR rep_origin 1086..1541 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1173..1192) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1383..1404 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1558..1577) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1677..1699 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1737..1755) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1823..1927 /gene="bla" /label=AmpR promoter CDS 1928..2788 /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(2146..2165) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2959..3547 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3448..3467 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtggagtt ccgcgttaca taacttacgg taaatggccc 181 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 241 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 301 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga 361 cagtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg 421 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat 481 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 541 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc 601 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 661 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag 721 aagacaccgg aattcggtac caccggtgct ctagacatat ggatatctga ctagtctcga 781 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 841 agcatttttt tcactggacc gagcggccgc aggaacccct agtgatggag ttggccactc 901 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg 961 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga 1021 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc 1081 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 1141 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct 1201 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 1261 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag 1321 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 1381 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga 1441 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa 1501 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac 1561 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc 1621 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg 1681 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct 1741 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg 1801 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc 1861 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag 1921 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg 1981 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt 2041 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt 2101 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt 2161 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa 2221 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag 2281 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac 2341 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac 2401 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac 2461 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac 2521 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact 2581 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg 2641 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt 2701 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat 2761 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta 2821 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa 2881 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 2941 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 3001 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 3061 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc 3121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 3181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 3241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 3301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 3361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 3421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 3481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 3541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 3601 tcacatgt //