LOCUS Exported 7118 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pAAV-Ef1a-FAS-NpHR3.0-eYFP-WPRE-pA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7118) AUTHORS Saunders A, Johnson CA, Sabatini BL TITLE Novel recombinant adeno-associated viruses for Cre activated and inactivated transgene expression in neurons. JOURNAL Front Neural Circuits. 2012 Jul 27;6:47. doi: 10.3389/fncir.2012.00047. eCollection 2012. PUBMED 22866029 REFERENCE 2 (bases 1 to 7118) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7118 /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" CDS 2440..3153 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(2482..2503) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2743..2762) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3090..3111 /label=EGFP-C /note="EGFP, forward primer" misc_feature 3245..3833 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3298..3318) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3716..3727) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3865..4341 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4064..4083) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4381..4521 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4381..4510 /label=AAV2 ITR rep_origin 4596..5051 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4683..4702) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4893..4914 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5068..5087) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5187..5209 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5247..5265) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5333..5437 /gene="bla" /label=AmpR promoter CDS 5438..6298 /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(5656..5675) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6469..7057 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6958..6977 /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 ctctagagtc 1441 gactccggaa caacttcgta tatacctttc tatacgaagt tgtggcgcgc cgccaccatg 1501 acagagaccc tgcctcccgt gaccgagagt gccgtggccc ttcaagccga ggttacccaa 1561 agggagttgt tcgagttcgt gctgaacgac cctttgcttg caagcagtct ctatatcaac 1621 atcgcacttg caggactgag tatactgctg ttcgttttta tgacccgagg actcgatgat 1681 ccacgggcaa aacttattgc tgtgtcaacc atccttgtgc ctgtcgtcag cattgcctcc 1741 tacactggat tggcgagcgg cctgacaatt tccgttcttg aaatgccagc gggccatttt 1801 gcagaaggca gctcagtgat gctgggagga gaagaggtag atggtgtagt caccatgtgg 1861 ggacggtatc tcacctgggc actttccacg cccatgattc tcctcgctct gggtctcctg 1921 gccggaagca atgctacaaa gctcttcaca gctatcactt tcgatatcgc tatgtgcgtg 1981 actggccttg ccgcggccct gactacctcc tcccacctca tgagatggtt ctggtacgct 2041 atcagttgtg catgctttct ggtggtcttg tatatcctgc tggtggagtg ggcacaggac 2101 gccaaagccg cgggaaccgc tgacatgttc aataccctga agctgttgac agtagtgatg 2161 tggctggggt atccaattgt gtgggctctt ggagtcgagg gtatcgcggt gttgcccgtt 2221 ggggtgacga gctggggata ttctttcctg gatatcgtgg caaagtacat tttcgcattc 2281 ttgctcctga actatctgac gtcaaacgaa tctgtcgtgt ccggcagcat tttggatgtt 2341 ccatctgctt ctgggacccc ggctgatgat gcggccgcca agagcaggat caccagcgag 2401 ggcgagtaca tccccctgga ccagatcgac atcaacgtgg tgagcaaggg cgaggagctg 2461 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc 2521 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc 2581 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc 2641 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc 2701 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag 2761 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc 2821 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc 2881 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc 2941 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc 3001 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg 3061 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 3121 gggatcactc tcggcatgga cgagctgtac aagttctgct acgagaacga ggtgtaatga 3181 gctagcacaa cttcgtatat acctttctat acgaagttgt gaattcgata tcaagcttat 3241 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 3301 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 3361 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 3421 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 3481 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 3541 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 3601 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 3661 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 3721 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 3781 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata 3841 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc 3901 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc 3961 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg 4021 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc 4081 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt 4141 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa 4201 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc 4261 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc 4321 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc 4381 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 4441 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 4501 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg 4561 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag 4621 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 4681 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 4741 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 4801 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 4861 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 4921 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta 4981 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac 5041 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca 5101 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc 5161 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc 5221 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt 5281 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac 5341 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 5401 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt 5461 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct 5521 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga 5581 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag 5641 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca 5701 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga 5761 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag 5821 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc 5881 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa 5941 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt 6001 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg 6061 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt 6121 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg 6181 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat 6241 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact 6301 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa 6361 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 6421 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 6481 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 6541 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 6601 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt 6661 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 6721 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 6781 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 6841 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 6901 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 6961 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 7021 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 7081 acggttcctg gccttttgct ggccttttgc tcacatgt //