LOCUS Exported 6605 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses Flpo in Mammalian Cells. ACCESSION . VERSION . KEYWORDS pAAV-EF1a-Flpo SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6605) 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 6605) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6605 /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" misc_feature 2732..3320 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2785..2805) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3203..3214) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3352..3828 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3551..3570) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3868..4008 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3868..3997 /label=AAV2 ITR rep_origin 4083..4538 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4170..4189) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4380..4401 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4555..4574) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4674..4696 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4734..4752) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4820..4924 /gene="bla" /label=AmpR promoter CDS 4925..5785 /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(5143..5162) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5956..6544 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6445..6464 /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 ccaccatgag 1441 ccagttcgac atcctgtgca agaccccccc caaggtgctg gtgcggcagt tcgtggagag 1501 attcgagagg cccagcggcg agaagatcgc cagctgtgcc gccgagctga cctacctgtg 1561 ctggatgatc acccacaacg gcaccgccat caagagggcc accttcatga gctacaacac 1621 catcatcagc aacagcctga gcttcgacat cgtgaacaag agcctgcagt tcaagtacaa 1681 gacccagaag gccaccatcc tggaggccag cctgaagaag ctgatccccg cctgggagtt 1741 caccatcatc ccttacaacg gccagaagca ccagagcgac atcaccgaca tcgtgtccag 1801 cctgcagctg cagttcgaga gcagcgagga ggccgacaag ggcaacagcc acagcaagaa 1861 gatgctgaag gccctgctgt ccgagggcga gagcatctgg gagatcaccg agaagatcct 1921 gaacagcttc gagtacacca gcaggttcac caagaccaag accctgtacc agttcctgtt 1981 cctggccaca ttcatcaact gcggcaggtt cagcgacatc aagaacgtgg accccaagag 2041 cttcaagctg gtgcagaaca agtacctggg cgtgatcatt cagtgcctgg tgaccgagac 2101 caagacaagc gtgtccaggc acatctactt tttcagcgcc agaggcagga tcgaccccct 2161 ggtgtacctg gacgagttcc tgaggaacag cgagcccgtg ctgaagagag tgaacaggac 2221 cggcaacagc agcagcaaca agcaggagta ccagctgctg aaggacaacc tggtgcgcag 2281 ctacaacaag gccctgaaga agaacgcccc ctaccccatc ttcgctatca agaacggccc 2341 taagagccac atcggcaggc acctgatgac cagctttctg agcatgaagg gcctgaccga 2401 gctgacaaac gtggtgggca actggagcga caagagggcc tccgccgtgg ccaggaccac 2461 ctacacccac cagatcaccg ccatccccga ccactacttc gccctggtgt ccaggtacta 2521 cgcctacgac cccatcagca aggagatgat cgccctgaag gacgagacca accccatcga 2581 ggagtggcag cacatcgagc agctgaaggg cagcgccgag ggcagcatca gataccccgc 2641 ctggaacggc atcatcagcc aggaggtgct ggactacctg agcagctaca tcaacaggcg 2701 gatctgagaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga 2761 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt 2821 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa 2881 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt 2941 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct 3001 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg 3061 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc 3121 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg 3181 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct 3241 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc 3301 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat 3361 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc 3421 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat 3481 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg 3541 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca 3601 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc 3661 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc 3721 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc 3781 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag 3841 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct 3901 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct 3961 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc 4021 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata 4081 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 4141 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 4201 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 4261 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg 4321 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 4381 tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt 4441 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 4501 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat 4561 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc 4621 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag 4681 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt 4741 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg 4801 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa 4861 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 4921 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct 4981 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg 5041 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg 5101 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt 5161 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga 5221 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga 5281 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac 5341 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg 5401 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac 5461 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct 5521 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct 5581 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg 5641 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat 5701 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg 5761 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat 5821 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 5881 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 5941 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 6001 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 6061 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta 6121 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 6181 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 6241 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 6301 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 6361 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 6421 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 6481 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 6541 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca 6601 catgt //