LOCUS       Exported                7148 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 Grm2. Synapsin promoter for neuronal expression..
ACCESSION   .
VERSION     .
KEYWORDS    LITE2.0_pAAV_hSyn_TALE(Grm2)-GS-CIB1(NLS*,∆318-334)_WPRE_bGHpA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7148)
  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 7148)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7148
                     /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"
     misc_feature    3540..4128
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3593..3613)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4011..4022)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4170..4187)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4176..4383
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4412..4552
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4412..4541
                     /label=AAV2 ITR
     rep_origin      4626..5081
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4713..4732)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4923..4944
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5098..5117)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5217..5239
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5277..5295)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5363..5467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5468..6328
                     /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(5686..5705)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6499..7087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6988..7007
                     /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 tagcggcggc ggcggcagcg
     2581 gcggcggcgg cagcaacggc gcgattggtg gggatttgct gcttaacttt cccgacatgt
     2641 ccgtgttgga acgtcagcgc gcacatttga agtatcttaa ccccaccttc gactccccgt
     2701 tggccgggtt ctttgcggac tcatctatga ttacgggagg ggaaatggac agctacctct
     2761 caacggccgg attgaatctt ccgatgatgt atggagaaac cactgtagaa ggcgactcgc
     2821 gactctcgat ttcgcctgaa acgacgctgg gaacagggaa cttcaaggcg gcgaaattcg
     2881 acacggagac aaaagattgc aacgaagcgg cgaagaaaat gaccatgaat cgcgatgatc
     2941 tggtagagga gggagaggag gaaaagtcga agattactga acagaacaat gggtctacca
     3001 aaagtatcaa aaagatgaag cacaaagcta agaaagaaga gaacaatttc agcaatgaca
     3061 gcagtaaagt cacaaaagaa ctggagaaaa cggattacat tcacgtgagg gcgcgacgag
     3121 ggcaggctac agattcacat tcaattgcgg agagagtacg gagagagaaa atctcagaaa
     3181 ggatgaagtt cctccaagac cttgtgccag gttgtgacaa gatcacaggc aaagcaggaa
     3241 tgctggatga gatcatcaac tacgtccaat cgttgcaaag acaaattgag tttctctcga
     3301 tgaaactggc catcgtgaat cctagaccgg atttcgacat ggatgacatc tttgcgaaag
     3361 aagtggcatc cactcccatg acggttgtgc cctcaccgga gatggtcttg tctggttaca
     3421 gccacgaaat ggtgcattcg ggttattcaa gcgagatggt caattcggga taccttcacg
     3481 tcaatcccat gcagcaggtg aatacttcca gttaagaatt cgatatcaag cttatcgata
     3541 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3601 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3661 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3721 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3781 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3841 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3901 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3961 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4021 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4081 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     4141 cgctgctcga gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     4201 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     4261 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4321 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     4381 ggaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     4441 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4501 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     4561 atgcggtatt ttctccttac gcatctgtgc ggtattcaca ccgcatacgt caaagcaacc
     4621 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4681 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4741 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4801 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     4861 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4921 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga
     4981 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5041 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     5101 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5161 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5221 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5281 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5341 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5401 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5461 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5521 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5581 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5641 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5701 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5761 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5821 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5881 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5941 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6001 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6061 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6121 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6181 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6241 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6301 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6361 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6421 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6481 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6541 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6601 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     6661 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6721 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6781 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6841 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6901 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6961 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7021 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7081 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7141 tcacatgt
//
