LOCUS Exported 8909 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION LITE2.0 TALE-CIB1. Changes from LITE1.0: glycine-serine linker, mutated endogenous NLS, aa318-334 deletion. This particular TALE targeted to Neurog2. EF1-alpha promoter for ubiquitous mammalian exp.. ACCESSION . VERSION . KEYWORDS LITE2.0 EF1a_TALE(Neurog2)-GS-CIB1(NLS*,∆318-334)_WPRE_hGHpA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8909) 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 8909) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8909 /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 237..1414 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 468..1405 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1362..1382 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 1430..1456 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" misc_feature 5036..5624 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5089..5109) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5507..5518) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 5656..6132 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(5855..5874) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 6172..6312 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 6172..6301 /label=AAV2 ITR rep_origin 6387..6842 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6474..6493) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6684..6705 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6859..6878) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6978..7000 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(7038..7056) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 7124..7228 /gene="bla" /label=AmpR promoter CDS 7229..8089 /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(7447..7466) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8260..8848 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8749..8768 /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 aagctttgca aagatggata aagttttaaa 181 cagagaggaa tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct 241 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 301 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 361 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 421 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt 481 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt 541 ccactggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag 601 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg 661 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga 721 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga 781 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg 841 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg 901 gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct 961 cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc 1021 gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg 1081 gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc 1141 agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt 1201 ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt 1261 tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc 1321 cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt 1381 tcaaagtttt tttcttccat ttcaggtgtc gtgaggatcc gccaccatgt acccatacga 1441 tgttccagat tacgcttcgc ggacccggct cccttcccca cccgcaccca gcccagcgtt 1501 ttcggccgac tcgttctcag acctgcttag gcagttcgac ccctcactgt ttaacacatc 1561 gttgttcgac tcccttcctc cgtttggggc gcaccatacg gaggcggcca ccggggagtg 1621 ggatgaggtg cagtcgggat tgagagctgc ggatgcacca cccccaacca tgcgggtggc 1681 cgtcaccgct gcccgaccgc cgagggcgaa gcccgcacca aggcggaggg cagcgcaacc 1741 gtccgacgca agccccgcag cgcaagtaga tttgagaact ttgggatatt cacagcagca 1801 gcaggaaaag atcaagccca aagtgaggtc gacagtcgcg cagcatcacg aagcgctggt 1861 gggtcatggg tttacacatg cccacatcgt agccttgtcg cagcaccctg cagcccttgg 1921 cacggtcgcc gtcaagtacc aggacatgat tgcggcgttg ccggaagcca cacatgaggc 1981 gatcgtcggt gtggggaaac agtggagcgg agcccgagcg cttgaggccc tgttgacggt 2041 cgcgggagag ctgagagggc ctccccttca gctggacacg ggccagttgc tgaagatcgc 2101 gaagcgggga ggagtcacgg cggtcgaggc ggtgcacgcg tggcgcaatg cgctcacggg 2161 agcacccctc aacctgaccc cagagcaggt cgtggcaatt gcgagccata acgggggaaa 2221 gcaggcactc gaaaccgtcc agaggttgct gcctgtgctg tgccaagcgc acggacttac 2281 gccagagcag gtcgtggcaa ttgcgagcaa catcggggga aagcaggcac tcgaaaccgt 2341 ccagaggttg ctgcctgtgc tgtgccaagc gcacggacta accccagagc aggtcgtggc 2401 aattgcgagc aacatcgggg gaaagcaggc actcgaaacc gtccagaggt tgctgcctgt 2461 gctgtgccaa gcgcacgggt tgaccccaga gcaggtcgtg gcaattgcga gcaacggagg 2521 gggaaagcag gcactcgaaa ccgtccagag gttgctgcct gtgctgtgcc aagcgcacgg 2581 cttaacccca gagcaggtcg tggcaattgc gagccataac gggggaaagc aggcactcga 2641 aaccgtccag aggttgctgc ctgtgctgtg ccaagcgcac ggcctgaccc cagagcaggt 2701 cgtggcaatt gcgagcaaca tcgggggaaa gcaggcactc gaaaccgtcc agaggttgct 2761 gcctgtgctg tgccaagcgc acggactgac accagagcag gtcgtggcaa ttgcgagcaa 2821 cggaggggga aagcaggcac tcgaaaccgt ccagaggttg ctgcctgtgc tgtgccaagc 2881 gcacggactc actccagagc aggtcgtggc aattgcgagc cataacgggg gaaagcaggc 2941 actcgaaacc gtccagaggt tgctgcctgt gctgtgccaa gcgcacggcc tcaccccaga 3001 gcaggtcgtg gcaattgcga gcaacatcgg gggaaagcag gcactcgaaa ccgtccagag 3061 gttgctgcct gtgctgtgcc aagcgcacgg attaacccca gagcaggtcg tggcaattgc 3121 gagcaacgga gggggaaagc aggcactcga aaccgtccag aggttgctgc ctgtgctgtg 3181 ccaagcgcac ggacttaccc cagagcaggt cgtggcaatt gcgagcaaca tcgggggaaa 3241 gcaggcactc gaaaccgtcc agaggttgct gcctgtgctg tgccaagcgc acgggctcac 3301 cccagagcag gtcgtggcaa ttgcgagcaa catcggggga aagcaggcac tcgaaaccgt 3361 ccagaggttg ctgcctgtgc tgtgccaagc gcacggtctt accccagagc aggtcgtggc 3421 aattgcgagc aacggagggg gaaagcaggc actcgaaacc gtccagaggt tgctgcctgt 3481 gctgtgccaa gcgcacggcc tcaccccaga gcaggtcgtg gcaattgcga gcaacatcgg 3541 gggaaagcag gcactcgaaa ccgtccagag gttgctgcct gtgctgtgcc aagcgcatgg 3601 gctcacccca gagcaggtcg tggcaattgc gagccatgac gggggaaagc aggcactcga 3661 aaccgtccag aggttgctgc ctgtgctgtg ccaagcgcac ggtctaaccc cagagcaggt 3721 cgtggcaatt gcgagccata acgggggaaa gcaggcactc gaaaccgtcc agaggttgct 3781 gcctgtgctg tgccaagcgc acggactcac gcctgagcag gtagtggcta ttgcatccaa 3841 taacgggggc agacccgcac tggagtcaat cgtggcccag ctttcgaggc cggaccccgc 3901 gctggccgca ctcactaatg atcatcttgt agcgctggcc tgcctcggcg gacgacccgc 3961 cttggatgcg gtgaagaagg ggctcccgca cgcgcctgca ttgattaagc ggaccaacag 4021 aaggattccc gagaggacat cacatcgagt ggcagctagc ggcggcggcg gcagcggcgg 4081 cggcggcagc aacggcgcga ttggtgggga tttgctgctt aactttcccg acatgtccgt 4141 gttggaacgt cagcgcgcac atttgaagta tcttaacccc accttcgact ccccgttggc 4201 cgggttcttt gcggactcat ctatgattac gggaggggaa atggacagct acctctcaac 4261 ggccggattg aatcttccga tgatgtatgg agaaaccact gtagaaggcg actcgcgact 4321 ctcgatttcg cctgaaacga cgctgggaac agggaacttc aaggcggcga aattcgacac 4381 ggagacaaaa gattgcaacg aagcggcgaa gaaaatgacc atgaatcgcg atgatctggt 4441 agaggaggga gaggaggaaa agtcgaagat tactgaacag aacaatgggt ctaccaaaag 4501 tatcaaaaag atgaagcaca aagctaagaa agaagagaac aatttcagca atgacagcag 4561 taaagtcaca aaagaactgg agaaaacgga ttacattcac gtgagggcgc gacgagggca 4621 ggctacagat tcacattcaa ttgcggagag agtacggaga gagaaaatct cagaaaggat 4681 gaagttcctc caagaccttg tgccaggttg tgacaagatc acaggcaaag caggaatgct 4741 ggatgagatc atcaactacg tccaatcgtt gcaaagacaa attgagtttc tctcgatgaa 4801 actggccatc gtgaatccta gaccggattt cgacatggat gacatctttg cgaaagaagt 4861 ggcatccact cccatgacgg ttgtgccctc accggagatg gtcttgtctg gttacagcca 4921 cgaaatggtg cattcgggtt attcaagcga gatggtcaat tcgggatacc ttcacgtcaa 4981 tcccatgcag caggtgaata cttccagtta agaattcgat atcaagctta tcgataatca 5041 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 5101 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 5161 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 5221 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 5281 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 5341 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 5401 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta 5461 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 5521 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 5581 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct 5641 gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg 5701 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg 5761 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg 5821 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca 5881 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct 5941 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt 6001 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag 6061 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc 6121 ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc 6181 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac 6241 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca 6301 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 6361 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg 6421 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 6481 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 6541 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 6601 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 6661 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 6721 tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 6781 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat 6841 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca 6901 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag 6961 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa 7021 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat 7081 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 7141 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 7201 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 7261 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 7321 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 7381 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 7441 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg 7501 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 7561 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 7621 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 7681 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 7741 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 7801 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 7861 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 7921 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 7981 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 8041 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 8101 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 8161 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 8221 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 8281 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 8341 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 8401 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 8461 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 8521 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 8581 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 8641 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 8701 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 8761 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 8821 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 8881 ggccttttgc tggccttttg ctcacatgt //