LOCUS Exported 7236 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION epiLITE / LITE1.0 TALE-CIB1. CIB1 binds to blue-light-activated CRY2PHR. Particular TALE targeted to Grm2 promoter. Synapsin promoter for neuronal expression.. ACCESSION . VERSION . KEYWORDS LITE1.0_pAAV_hSyn_TALE(Grm2)-NLS-CIB1_2A_GFP_WPRE_bGHpA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7236) AUTHORS Konermann S, Brigham MD, Trevino A, Hsu PD, Heidenreich M, Le Cong, Platt RJ, Scott DA, Church GM, Zhang F TITLE Optical control of mammalian endogenous transcription and epigenetic states. JOURNAL Nature. 2013 Jul 23. doi: 10.1038/nature12466. PUBMED 23877069 REFERENCE 2 (bases 1 to 7236) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7236 /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 181..628 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 651..677 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2568..2588 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" misc_feature 3627..4215 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3680..3700) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4098..4109) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4257..4274) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4263..4470 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 4499..4639 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4499..4628 /label=AAV2 ITR rep_origin 4714..5169 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4801..4820) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5011..5032 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5186..5205) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5305..5327 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5365..5383) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5451..5555 /gene="bla" /label=AmpR promoter CDS 5556..6416 /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(5774..5793) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6587..7175 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7076..7095 /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 gcgttgtaca gtgtctagac tgcagagggc cctgcgtatg 181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga 241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga 301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg 361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc 421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg 481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg 541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg 601 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggatc cgccaccatg tacccatacg 661 atgttccaga ttacgctgta gatttgagaa ctttgggata ttcacagcag cagcaggaaa 721 agatcaagcc caaagtgagg tcgacagtcg cgcagcatca cgaagcgctg gtgggtcatg 781 ggtttacaca tgcccacatc gtagccttgt cgcagcaccc tgcagccctt ggcacggtcg 841 ccgtcaagta ccaggacatg attgcggcgt tgccggaagc cacacatgag gcgatcgtcg 901 gtgtggggaa acagtggagc ggagcccgag cgcttgaggc cctgttgacg gtcgcgggag 961 agctgagagg gcctcccctt cagctggaca cgggccagtt gctgaagatc gcgaagcggg 1021 gaggagtcac ggcggtcgag gcggtgcacg cgtggcgcaa tgcgctcacg ggagcacccc 1081 tcaacctgac cccagagcag gtcgtggcga tcgcaagcca cgacggagga aagcaagcct 1141 tggaaacagt acagaggctg ttgcctgtgc tgtgccaagc gcacggactt accccagagc 1201 aggtcgtggc aatcgcctcc aacattggcg ggaaacaggc actcgagact gtccagcgcc 1261 tgcttcccgt gctgtgccaa gcgcacggcc ttaccccaga gcaggtcgtg gcaatcgcga 1321 gcaataacgg cggaaaacag gctttggaaa cggtgcagag gctccttcca gtgctgtgcc 1381 aagcgcacgg actaacccca gagcaggtcg tggcaatcgc ctccaacatt ggcgggaaac 1441 aggcactcga gactgtccag cgcctgcttc ccgtgctgtg ccaagcgcac gggctaaccc 1501 cagagcaggt cgtggcaatc gcgagcaata acggcggaaa acaggctttg gaaacggtgc 1561 agaggctcct tccagtgctg tgccaagcgc acggcctaac cccagagcag gtcgtggcga 1621 tcgcaagcca cgacggagga aagcaagcct tggaaacagt acagaggctg ttgcctgtgc 1681 tgtgccaagc gcacggactt acacccgaac aagtcgtggc aattgcgagc aacggagggg 1741 gaaagcaggc actcgaaacc gtccagaggt tgctgcctgt gctgtgccaa gcgcacggac 1801 ttaccccaga gcaggtcgtg gcaatcgcga gcaataacgg cggaaaacag gctttggaaa 1861 cggtgcagag gctccttcca gtgctgtgcc aagcgcacgg ccttacccca gagcaggtcg 1921 tggccattgc ctcgaatgga gggggcaaac aggcgttgga aaccgtacaa cgattgctgc 1981 cggtgctgtg ccaagcgcac ggactaaccc cagagcaggt cgtggcgatc gcaagccacg 2041 acggaggaaa gcaagccttg gaaacagtac agaggctgtt gcctgtgctg tgccaagcgc 2101 acgggctaac cccagagcag gtcgtggcga tcgcaagcca cgacggagga aagcaagcct 2161 tggaaacagt acagaggctg ttgcctgtgc tgtgccaagc gcacggccta accccagagc 2221 aggtcgtggc cattgcctcg aatggagggg gcaaacaggc gttggaaacc gtacaacgat 2281 tgctgccggt gctgtgccaa gcgcacggac tcacgcctga gcaggtagtg gctattgcat 2341 cccatgacgg gggcagaccc gcactggagt caatcgtggc ccagctttcg aggccggacc 2401 ccgcgctggc cgcactcact aatgatcatc ttgtagcgct ggcctgcctc ggcggacgac 2461 ccgccttgga tgcggtgaag aaggggctcc cgcacgcgcc tgcattgatt aagcggacca 2521 acagaaggat tcccgagagg acatcacatc gagtggcagc tagcagcccc aagaagaaga 2581 gaaaggtgga ggccagcaac ggcgcgattg gtggggattt gctgcttaac tttcccgaca 2641 tgtccgtgtt ggaacgtcag cgcgcacatt tgaagtatct taaccccacc ttcgactccc 2701 cgttggccgg gttctttgcg gactcatcta tgattacggg aggggaaatg gacagctacc 2761 tctcaacggc cggattgaat cttccgatga tgtatggaga aaccactgta gaaggcgact 2821 cgcgactctc gatttcgcct gaaacgacgc tgggaacagg gaacttcaag aaacggaaat 2881 tcgacacgga gacaaaagat tgcaacgaaa agaagaagaa aatgaccatg aatcgcgatg 2941 atctggtaga ggagggagag gaggaaaagt cgaagattac tgaacagaac aatgggtcta 3001 ccaaaagtat caaaaagatg aagcacaaag ctaagaaaga agagaacaat ttcagcaatg 3061 acagcagtaa agtcacaaaa gaactggaga aaacggatta cattcacgtg agggcgcgac 3121 gagggcaggc tacagattca cattcaattg cggagagagt acggagagag aaaatctcag 3181 aaaggatgaa gttcctccaa gaccttgtgc caggttgtga caagatcaca ggcaaagcag 3241 gaatgctgga tgagatcatc aactacgtcc aatcgttgca aagacaaatt gagtttctct 3301 cgatgaaact ggccatcgtg aatcctagac cggatttcga catggatgac atctttgcga 3361 aagaagtggc atccactccc atgacggttg tgccctcacc ggagatggtc ttgtctggtt 3421 acagccacga aatggtgcat tcgggttatt caagcgagat ggtcaattcg ggataccttc 3481 acgtcaatcc catgcagcag gtgaatactt ccagtgatcc actctcctgc tttaacaacg 3541 gcgaggcccc ttcgatgtgg gactcccacg tacagaatct ctatggaaat ctcggagtct 3601 aagaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat 3661 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc 3721 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct 3781 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca 3841 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt 3901 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg 3961 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga 4021 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt 4081 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc 4141 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt 4201 gggccgcctc cccgcatcga taccgagcgc tgctcgagct agagctcgct gatcagcctc 4261 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac 4321 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg 4381 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga 4441 ttgggaagac aatagcaggc atgctgggga ggtaaccacg tgcggaccga gcggccgcag 4501 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc 4561 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga 4621 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt 4681 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg 4741 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 4801 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 4861 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 4921 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 4981 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac 5041 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt 5101 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt 5161 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc 5221 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg 5281 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat 5341 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca 5401 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc 5461 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 5521 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg 5581 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg 5641 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc 5701 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca 5761 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac 5821 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa 5881 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg 5941 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt 6001 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg 6061 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc 6121 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga 6181 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta 6241 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc 6301 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg 6361 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt 6421 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa 6481 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt 6541 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 6601 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 6661 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 6721 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 6781 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 6841 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 6901 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 6961 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 7021 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 7081 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 7141 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 7201 ggttcctggc cttttgctgg ccttttgctc acatgt //