LOCUS Exported 7636 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses Dre in Mammalian Cells. ACCESSION . VERSION . KEYWORDS pAAV-EF1a-mCherry-IRES-Dre SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7636) 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 7636) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7636 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 231..1409 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 462..1400 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1357..1377 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 1422..2129 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE GRHSTGGMDELYK" primer_bind complement(1564..1582) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1832..1851 /label=mCherry-F /note="mCherry, forward primer" misc_feature 2146..2698 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(2290..2307) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 2517..2536 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" misc_feature 3763..4351 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3816..3836) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4234..4245) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4383..4859 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4582..4601) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4899..5039 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4899..5028 /label=AAV2 ITR rep_origin 5114..5569 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5201..5220) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5411..5432 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5586..5605) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5705..5727 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5765..5783) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5851..5955 /gene="bla" /label=AmpR promoter CDS 5956..6816 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGNQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6174..6193) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6987..7575 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7476..7495 /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga 181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg 241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg 301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt 361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg 421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt 481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct 541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga 601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct 661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt 721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc 781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg 841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac 901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc 961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag 1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct 1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg 1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga 1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc 1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg 1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa 1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc cgtgagcaag ggcgaggagg 1441 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga 1501 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga 1561 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc 1621 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact 1681 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg 1741 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga 1801 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct 1861 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc 1921 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg 1981 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct 2041 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca 2101 ccggcggcat ggacgagctg tacaagtaag gcgcgccccc ccctaacgtt actggccgaa 2161 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt 2221 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg 2281 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc 2341 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc 2401 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa 2461 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc 2521 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg 2581 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac 2641 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg 2701 gccacaacca tgtcagagct tataatttca ggtagtagtg gaggatttct cagaaacatt 2761 ggcaaggagt accaagaagc tgccgaaaat ttcatgaggt ttatgaacga ccagggcgca 2821 tacgctccaa atactttgag agacctccga ctcgtgttcc acagctgggc tcgatggtgc 2881 cacgccaggc agcttgcatg gtttccaata tctccagaaa tggctcgcga gtactttttg 2941 caactccacg acgctgatct ggcatctacc accatcgaca agcactatgc aatgctgaat 3001 atgctgctgt cccactgtgg actgcctcca ctgtcagatg acaagtccgt gagcctggcg 3061 atgcggagaa ttagaagaga ggcggccact gaaaaggggg aacggacagg acaggcgata 3121 cctctccgct gggacgacct gaaactgctg gacgtactcc tgagtcggag cgagcgcttg 3181 gtcgatcttc ggaaccgcgc cttcctcttc gtggcctata ataccctgat gcggatgagt 3241 gagatcagca gaatccgagt tggggatctg gatcagaccg gggacactgt gactttgcat 3301 atctcccata ctaagactat caccaccgct gccggcttgg acaaggttct ctctaggcgc 3361 acgaccgctg tcctgaatga ctggcttgat gtatctggcc tgcgagaaca tcccgacgcc 3421 gtgctgttcc cccctattca ccggtctaac aaggcaagaa tcacgacaac gccgctgact 3481 gcacccgcca tggagaagat cttttccgat gcttgggtgc tccttaacaa aagggatgca 3541 acacctaaca aaggtagata caggacatgg accgggcata gcgcgcgggt gggggccgcg 3601 attgatatgg ccgagaaaca ggtcagcatg gtggagatca tgcaggaggg tacatggaaa 3661 aaaccggaga cactgatgag atatcttagg agggggggcg tcagcgttgg agcaaatagc 3721 cgcctgatgg acagttgaga attcgatatc aagcttatcg ataatcaacc tctggattac 3781 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 3841 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 3901 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 3961 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 4021 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 4081 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 4141 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg 4201 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 4261 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 4321 agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct 4381 acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc 4441 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt 4501 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga 4561 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt 4621 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt 4681 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac 4741 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac 4801 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc 4861 tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt 4921 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg 4981 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg 5041 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca 5101 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 5161 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct 5221 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 5281 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt 5341 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 5401 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat 5461 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt 5521 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact 5581 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc 5641 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 5701 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga 5761 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag 5821 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 5881 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 5941 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 6001 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 6061 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 6121 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 6181 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 6241 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 6301 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 6361 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 6421 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 6481 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 6541 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 6601 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 6661 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 6721 atcgtagtta tctacacgac ggggaatcag gcaactatgg atgaacgaaa tagacagatc 6781 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 6841 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 6901 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 6961 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 7021 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 7081 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 7141 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 7201 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 7261 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 7321 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 7381 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 7441 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 7501 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 7561 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 7621 ccttttgctc acatgt //