LOCUS Exported 5843 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cre-on/Flp-on EYFP under the Synapsin promoter. ACCESSION . VERSION . KEYWORDS pAAV-hSyn Con/Fon EYFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5843) AUTHORS Fenno LE, Mattis J, Ramakrishnan C, Hyun M, Lee SY, He M, Tucciarone J, Selimbeyoglu A, Berndt A, Grosenick L, Zalocusky KA, Bernstein H, Swanson H, Perry C, Diester I, Boyce FM, Bass CE, Neve R, Huang ZJ, Deisseroth K TITLE Targeting cells with single vectors using multiple-feature Boolean logic. JOURNAL Nat Methods. 2014 Jul;11(7):763-72. doi: 10.1038/nmeth.2996. Epub 2014 Jun 8. PUBMED 24908100 REFERENCE 2 (bases 1 to 5843) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5843 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 249..282 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 333..366 /label=loxN /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTATACCT). loxN sites are compatible with each other, but incompatible with loxP or lox2272 sites." primer_bind 479..500 /label=EGFP-N /note="EGFP, reverse primer" regulatory 542..551 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(556..589) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(640..673) /label=loxN /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTATACCT). loxN sites are compatible with each other, but incompatible with loxP or lox2272 sites." CDS complement(809..1060) /codon_start=1 /product="C-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /label=VC155 /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK" primer_bind complement(851..872) /label=EGFP-C /note="EGFP, forward primer" primer_bind 1200..1219 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 1583..2171 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1636..1656) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2054..2065) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2203..2679 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2402..2421) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 2719..2859 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2719..2848 /label=AAV2 ITR rep_origin 2934..3389 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3021..3040) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3231..3252 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3406..3425) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3525..3547 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3585..3603) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3671..3775 /gene="bla" /label=AmpR promoter CDS 3776..4636 /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(3994..4013) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4807..5395 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5296..5315 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 5457..5586 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 5631..235 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" ORIGIN 1 ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc 61 ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc 121 gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc 181 gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag cgcagtcgag 241 aaggatccat aacttcgtat aggatacttt atacgaagtt atgtagctgg agcaatttga 301 aaagattgtt gtattacctc tagctgcgct taataacttc gtatagtata ccttatacga 361 agttatacta gtcctcctgt actcaccttg ccggtggtgc agatgaactt cagggtcagc 421 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg 481 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc 541 accatggtgg ctagcataac ttcgtataaa gtatcctata cgaagttatt tgccttaacc 601 cagaaattat cactgttatt ctttagatag gtgcaaagaa taacttcgta taaggtatac 661 tatacgaagt tatggtaccg aagttcctat tcttcaaata gtataggaac ttcaattgtt 721 attggtgcaa agaatcactt ccttaagtct ttatactgcc caggaagttc ctattcttca 781 aaaggtatag gaacttcggc gcgccttact tgtacagctc gtccatgccg agagtgatcc 841 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc 901 tcagggcgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga 961 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc 1021 ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt 1081 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga 1141 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg 1201 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca 1261 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca 1321 ggccgtagcc gaaggtggtc acgagggtgg gccagggcac gggcagctgt gggacgacat 1381 gacttaacca gttaaccatg ttctctgaac atgagaaggg ctggccactc tccagagctc 1441 gaagttccta tactatttga agaataggaa cttcttgcct taacccagaa attatcactg 1501 ttattcttta gaatggtgca aagagaagtt cctatacctt ttgaagaata ggaacttcga 1561 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac 1621 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 1681 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 1741 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 1801 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 1861 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 1921 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc 1981 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt 2041 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 2101 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc 2161 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac 2221 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc 2281 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg 2341 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg 2401 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc 2461 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc 2521 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc 2581 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg 2641 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg 2701 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg 2761 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg 2821 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc 2881 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc 2941 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 3001 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 3061 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 3121 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc 3181 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 3241 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat 3301 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 3361 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga 3421 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc 3481 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg 3541 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct 3601 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg 3661 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc 3721 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag 3781 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt 3841 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt 3901 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga 3961 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat 4021 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga 4081 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag 4141 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg 4201 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg 4261 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt 4321 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg 4381 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc 4441 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg 4501 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac 4561 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact 4621 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa 4681 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa 4741 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg 4801 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc 4861 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac 4921 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca 4981 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt 5041 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc 5101 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg 5161 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc 5221 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac 5281 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct 5341 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc 5401 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg 5461 caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc 5521 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg 5581 gttcctgcgg ccgcacgcgt gtgtctagac tgcagagggc cctgcgtatg agtgcaagtg 5641 ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca 5701 ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg 5761 ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct 5821 tcgcccccgc ctggcggcgc gcg //