LOCUS Exported 6085 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION One half component of the optimized Cre recombinase dependent on GFP (CRE-DOG OPT) for rAAV production and expression in mammalian cells. ACCESSION . VERSION . KEYWORDS pAAV-EF1a-N-CretrcintG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6085) AUTHORS Tang JC, Rudolph S, Dhande OS, Abraira VE, Choi S, Lapan SW, Drew IR, Drokhlyansky E, Huberman AD, Regehr WG, Cepko CL TITLE Cell type-specific manipulation with GFP-dependent Cre recombinase. JOURNAL Nat Neurosci. 2015 Sep;18(9):1334-41. doi: 10.1038/nn.4081. Epub 2015 Aug 10. PUBMED 26258682 REFERENCE 2 (bases 1 to 6085) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6085 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR promoter 242..1419 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 473..1410 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1367..1387 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 1432..1441 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1468..1488 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" gap 1795..2184 /estimated_length=390 misc_feature 2212..2800 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2265..2285) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2683..2694) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2832..3308 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3031..3050) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3348..3488 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3348..3477 /label=AAV2 ITR rep_origin 3563..4018 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3650..3669) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3860..3881 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4035..4054) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4154..4176 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4214..4232) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4300..4404 /gene="bla" /label=AmpR promoter CDS 4405..5265 /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(4623..4642) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5436..6024 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5925..5944 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt 181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat 241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg 301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag 361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc 421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc 481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat 541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc cgccaccatg 1441 gataaagcgg aattaattcc cgagcctcca aaaaagaaga gaaaggtcga attgggtacc 1501 acctctgatg aagtcaggaa gaacctgatg gacatgttca gggacaggca ggccttctct 1561 gaacacacct ggaagatgct cctgtctgtg tgcagatcct gggctgcctg gtgcaagctg 1621 aacctcgtcg aaagctatcg caaggccagc aacaaggcct actttgagtg gaccatcgaa 1681 gccagggacc tctccctgct gggaagccac gtgcgcaagg ccacctacca gacctacgcc 1741 cccatcctgt acgctagcgg aggaggtggt ggaggtggag gtggaggtca attgnnnnnn 1801 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1861 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1921 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1981 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 2041 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 2101 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 2161 nnnnnnnnnn nnnnnnnnnn nnnntaagaa ttcgatatca agcttatcga taatcaacct 2221 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 2281 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 2341 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 2401 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 2461 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 2521 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 2581 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt 2641 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 2701 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 2761 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc 2821 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag 2881 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg 2941 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag 3001 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg 3061 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc 3121 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt 3181 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga 3241 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc 3301 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt 3361 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa 3421 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg 3481 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg 3541 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 3601 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 3661 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 3721 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg 3781 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 3841 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 3901 tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat 3961 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta 4021 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg 4081 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa 4141 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc 4201 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg 4261 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta 4321 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt 4381 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc 4441 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa 4501 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt 4561 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt 4621 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc 4681 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg 4741 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg 4801 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac 4861 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca 4921 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta 4981 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat 5041 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa 5101 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag 5161 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat 5221 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt 5281 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg 5341 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga 5401 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 5461 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 5521 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 5581 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 5641 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 5701 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 5761 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 5821 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 5881 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 5941 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 6001 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 6061 ttttgctggc cttttgctca catgt //