LOCUS Exported 6451 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses sCre in Mammalian Cells. ACCESSION . VERSION . KEYWORDS pAAV-EF1a-sCre SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6451) 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 6451) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6451 /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 2578..3166 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2631..2651) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3049..3060) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3198..3674 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3397..3416) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3714..3854 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3714..3843 /label=AAV2 ITR rep_origin 3929..4384 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4016..4035) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4226..4247 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4401..4420) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4520..4542 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4580..4598) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4666..4770 /gene="bla" /label=AmpR promoter CDS 4771..5631 /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(4989..5008) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5802..6390 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6291..6310 /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 agcctgctga ctaccaacaa ccattctgtg gcactgtcct acggcgaacc tccctctact 1501 ctgaacgatt ctctgaaaga ttcctaccag aggtctaccg acgagctgca ggcactgctg 1561 agtaagccac tggcacagct gaccgacgcc gataaactgc ggatcagaga aattacacag 1621 gctaagctga aacactttct ggataacggc catagaactc ggagagctaa tacctggaga 1681 gcactgatga gtaggtgggc caagttcgag tcatggtgcc tgaccaacaa tctgacaccc 1741 ctgcctgcaa cacccgaagt ggtcgccact tttatcgagt actatcaggc cagctcctac 1801 accacactga gccagtatgc ttgggcaatt aactccttcc acgtggaatg tggactgctg 1861 tctcccgtgt ctagtaagac agtccaggac aaacagaatg aaatccggat tgtgaagctg 1921 gagagcggag ggctggccca ggaacaggct actcctttta gactgcacca tctgcagatg 1981 ctgatcgagt cttacgggga aagtgagcgc ctgctggata aacgaaacct ggccctgctg 2041 aatatcgctt atgagtccct gctgcgggaa tctgagctgc tgagaattaa ggtcggccat 2101 ctgaaatcaa cattcgaagg ggactacgtg ctgagcgtcc cttatactaa gaccaacgat 2161 tccggggagg aagaggtggt caatatcact ccactgggat tcaagctgat ccagaggtac 2221 attcagggag ccggcctgac caaagaggac tatctgtttc agccaattgg ccgctcaaac 2281 aaggtgagcg tccaggccaa acccatgagc acacgaactg tggaccgggt cttcctgtgg 2341 gcttttgaat ccctggggat cgatcgacac tcagcttgga gcgggcattc cgcacgcatt 2401 ggagccgctc aggatctgct ggcagccgga tattctatcg cccagattca ggagaatggc 2461 aggtggaaaa gtcctatgat ggtgctgcgc tacggcaaag acatcaaggc aaaagaatcc 2521 gctatggcaa aaatgctggc agaaagaaga taagaattcg atatcaagct tatcgataat 2581 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 2641 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 2701 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 2761 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 2821 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 2881 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 2941 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc 3001 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 3061 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc 3121 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg 3181 ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc 3241 tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt 3301 tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg 3361 ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg 3421 cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc 3481 ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt 3541 ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc 3601 aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc 3661 ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc 3721 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg 3781 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg 3841 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc 3901 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg 3961 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 4021 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 4081 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 4141 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 4201 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 4261 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta 4321 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca 4381 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga 4441 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 4501 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 4561 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata 4621 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt 4681 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa 4741 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt 4801 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa 4861 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac 4921 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt 4981 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt 5041 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat 5101 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac 5161 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg 5221 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc 5281 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa 5341 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag 5401 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct 5461 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat 5521 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa 5581 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac 5641 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 5701 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 5761 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 5821 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 5881 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 5941 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 6001 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 6061 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 6121 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 6181 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 6241 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 6301 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 6361 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 6421 ctggcctttt gctggccttt tgctcacatg t //