LOCUS       sequence-72985-e        8909 bp    DNA     circular SYN 15-DEC-2025
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    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2013 Jul 23. doi: 10.1038/nature12466."
COMMENT     Sequence Label: LITE2.0
            EF1a_TALE(Neurog2)-GS-CIB1(NLS*,∆318-3...
FEATURES             Location/Qualifiers
     source          1..8909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(7229..7297,7298..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
//
